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WO2000009653A2 - Method of inhibiting cathepsin k - Google Patents

Method of inhibiting cathepsin k Download PDF

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Publication number
WO2000009653A2
WO2000009653A2 PCT/US1998/014634 US9814634W WO0009653A2 WO 2000009653 A2 WO2000009653 A2 WO 2000009653A2 US 9814634 W US9814634 W US 9814634W WO 0009653 A2 WO0009653 A2 WO 0009653A2
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WIPO (PCT)
Prior art keywords
lys
tyr
asn
leu
active site
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PCT/US1998/014634
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French (fr)
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WO2000009653A3 (en
Inventor
Ward W. Smith
Sherin Abdel-Meguid
Cheryl Janson
Judith Lalonde
Baoquang Zhao
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SmithKline Beecham Corp
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SmithKline Beecham Corp
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Priority to PCT/US1998/014634 priority Critical patent/WO2000009653A2/en
Priority to CA002301201A priority patent/CA2301201A1/en
Priority to JP2000565090A priority patent/JP2003527819A/en
Publication of WO2000009653A2 publication Critical patent/WO2000009653A2/en
Anticipated expiration legal-status Critical
Publication of WO2000009653A3 publication Critical patent/WO2000009653A3/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6402—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals
    • C12N9/6405—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from non-mammals not being snakes
    • C12N9/641—Cysteine endopeptidases (3.4.22)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A61P21/04—Drugs for disorders of the muscular or neuromuscular system for myasthenia gravis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
    • A61P33/08—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis for Pneumocystis carinii
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
    • C07K14/8139—Cysteine protease (E.C. 3.4.22) inhibitors, e.g. cystatin

Definitions

  • This invention relates to a method of inhibiting cathepsin K by the utilization of the physical and spacial characteristics that allow for the interaction of the propeptide with the active site of the enzyme. This interaction inhibits the activity of cathepsin K and this information is useful in the design of specific chemical compounds for treating diseases in which said inhibition is indicated, such as osteoporosis and periodontal disease.
  • This invention also relates to a novel crystalline structure of procathepsin K, the identification of the propeptide bound to the catalytic active site for this enzyme and methods enabling the design and selection of inhibitors of said active site.
  • Cathepsin K is a member of the family of enzymes which are part of the papain superfamily of cysteine proteases. Cathepsins B, H, L, N and S have been described in the literature. Recently, cathepsin K polypeptide and the cDNA encoding such polypeptide were disclosed in U.S. Patent No. 5,501,969 (called cathepsin O therein). Cathepsin K has been recently expressed, purified, and characterized. Bossard, M. J., et al., (1996) J. Biol. Chem. 271, 12517-12524; Drake, F.H., et al., (1996) J. Biol. Chem. 271, 12511-12516; Bromme, D., et al., (1996) J. Biol. Chem. Ill, 2126-2132.
  • Cathepsin K has been variously denoted as cathepsin O, cathepsin X or cathepsin O2 in the literature.
  • the designation cathepsin K is considered to be the more appropriate one (name assigned by Nomenclature Committee of the International Union of Biochemistry and Molecular Biology).
  • Cathepsins of the papain superfamily of cysteine proteases function in the normal physiological process of protein degradation in animals, including humans, e.g., in the degradation of connective tissue.
  • elevated levels of these enzymes in the body can result in pathological conditions leading to disease.
  • cathepsins have been implicated in various disease states, including but not limited to, infections by pneumocystis carinii, trypsanoma cruzi, trypsanoma brucei brucei, and Crithidia fusiculata; as well as in schistosomiasis malaria, tumor metastasis, metachromatic leukodystrophy, muscular dystrophy, amytrophy, and the like.
  • Bone is composed of a protein matrix in which spindle- or plate- shaped crystals of hydroxyapatite are incorporated.
  • Type I Collagen represents the major structural protein of bone comprising approximately 90% of the structural protein. The remaining 10% of matrix is composed of a number of non-collagenous proteins, including osteocalcin, proteoglycans, osteopontin, osteonectin, thrombospondin, fibronectin, and bone sialoprotein.
  • Skeletal bone undergoes remodeling at discrete foci throughout life. These foci, or remodeling units, undergo a cycle consisting of a bone resorption phase followed by a phase of bone replacement.
  • Bone resorption is carried out by osteoclasts, which are multinuclear cells of hematopoietic lineage.
  • the osteoclasts adhere to the bone surface and form a tight sealing zone, followed by extensive membrane ruffling on their apical (i.e., resorbing) surface.
  • the low pH of the compartment dissolves hydroxyapatite crystals at the bone surface, while the proteolytic enzymes digest the protein matrix. In this way, a resorption lacuna, or pit, is formed.
  • osteoblasts lay down a new protein matrix that is subsequently mineralized.
  • disease states such as osteoporosis and Paget's disease
  • the normal balance between bone resorption and formation is disrupted, and there is a net loss of bone at each cycle.
  • this leads to weakening of the bone and may result in increased fracture risk with minimal trauma.
  • cathepsin K may provide an effective treatment for diseases of excessive bone loss, including, but not limited to, osteoporosis, gingival diseases such as gingivitis and periodontitis, Paget's disease, hypercalcemia of malignancy, and metabolic bone disease.
  • Cathepsin K levels have also been demonstrated to be elevated in chondroclasts of osteoarthritic synovium.
  • selective inhibition of cathepsin K may also be useful for treating diseases of excessive cartilage or matrix degradation, including, but not limited to, osteoarthritis and rheumatoid arthritis.
  • Metastatic neoplastic cells also typically express high levels of proteolytic enzymes that degrade the surrounding matrix.
  • selective inhibition of cathepsin K may also be useful for treating certain neoplastic diseases.
  • the propeptide of cathepsin K exhibits characteristics that allow for the interaction of the propeptide with specific residues of the active site of cathepsin K and this information is useful in the design of specific chemical compounds as inhibitors of cathepsin K and would be useful for treating diseases in which inhibition of bone resorption is indicated, such as osteoporosis and periodontal disease.
  • this invention relates to the method of inhibiting cathepsin K using the crystal structure of procathepsin K hereinbelow defined.
  • the present invention provides a method for designing compounds to inhibit cathepsin K with certain structural, physical and spacial characteristics that allow for the interaction of said compounds with specific residues of the active site of the enzyme in a manner similar to the interaction of the propeptide with the enzyme active site. This interaction inhibits the activity of cathepsin K and, thus, treats diseases in which bone resorption is a factor.
  • the present invention provides a novel cysteine protease proenzyme in crystalline form, in particular this invention relates to a novel procathepsin K crystal.
  • the invention provides a novel protease proenzyme composition, specifically a novel procathepsin K composition characterized by a three dimensional catalytic site inhibited by the atoms of the propeptide residues listed in Table IV.
  • the invention provides a method for identifying inhibitors said method comprising the steps of: providing the coordinates of the proenzyme structure of the invention to a computerized modeling system; and identifying compounds which will bind to the structure in a manner similar to that of the propeptide.
  • Figure 1 is the amino acid sequence of procathepsin K aligned with the amino acid sequences of other cysteine protease proenzymes.
  • Figure 2 is a ribbon diagram of procathepsin K. The amino and carboxyl- termini are indicated by N and C. The drawing was produced using the program MOLSCRIPT [Kraulis, P., J. Appl. Crystallogr., 24, 946-950 (1991)].
  • Figure 3 is an illustration of the active site of procathepsin K.
  • Table I provides the three dimensional protein coordinates of the procathepsin K crystalline structure of the invention.
  • Table II provides a listing of the three atom angles between atoms of the propeptide and the protein for all propeptide atoms within 5 Angstroms of the protein.
  • Table III provides a listing of the distances between atoms of the propeptide and the protein for all propeptide atoms within 5 Angstroms of the protein.
  • Table IV provides the atoms of the amino acid residues of the catalytic site and the propeptide amino acid residues which interact with them.
  • the present invention provides a novel cysteine protease proenzyme crystalline structure, and methods of use of the crystalline form and active site interactions with the inhibitory propeptide to identify protease inhibitor compounds.
  • the present invention provides a method for inhibiting cathepsin K by designing compounds with certain structural, physical and spacial characteristics that allow for the interaction of said compounds with specific residues of the active site of the enzyme based on the structure of the proenzyme. This interaction inhibits the activity of cathepsin K and, thus, treats diseases in which bone resorption is a factor.
  • the present invention provides a novel cysteine protease proenzyme in crystalline form, in particular procathepsin K in crystalline form as defined by the positions in Table I herein.
  • the present invention provides a novel protease proenzyme composition, specifically a novel procathepsin K composition, characterized by a three dimensional catalytic site formed by the atoms of the amino acid residues listed in Table IV herein.
  • a key aspect of this invention provides a method of inhibiting cathepsin K which comprises administering to a mammal in need thereof a compound that mimics the interaction of the propeptide with the residues of the active site of procathepsin K.
  • this compound comprises: (i) an oxygen atom which interacts with a hydrogen atom donated by the indole nitrogen of tryptophan 283 wherein the distance between these two atoms is 2.7-3.5A;
  • the preferred compound consists of a hydrophobic group that interacts with said hydrophobic pocket wherein said hydrophobic group is an isobutyl group.
  • the method of the instant invention comprises compounds characterized by the interactions of the atoms of the procathepsin K active site and the atoms of the propeptide in the region of the acive site according to Table IV.
  • This invention also includes a composition comprising procathepsin K in crystalline form wherein the procathepsin K in crystalline form is characterized by structure coordinates according to Table I.
  • the composition has an active site cavity formed by the amino acids in Table IV wherein said active site is characterized by the distance between the atoms of Table II and the bond angles between interresidue atoms of Table III.
  • procathepsin K in another aspect of this invention is an isolated, properly folded procathepsin K molecule or fragment thereof having a conformation comprising a catalytically active site formed by the residues listed in Table IV, said active site defined by the protein coordinates of Table I and a peptide, peptidomimetic or synthetic molecule which binds with the active site cavity of procathepsin K.
  • this invention provides for a method of identifying an inhibitor compound capable of binding to, and inhibiting the proteolytic activity of, cathepsin K, said method comprising: introducing into a suitable computer program information defining an active site conformation of a procathepsin K molecule comprising a catalytically active site formed by the residues listed in Table IV, said active site defined by the protein coordinates of Table I, wherein said program displays the three-dimensional structure thereof; creating a three dimensional representation of the active site cavity in said computer program; displaying and superimposing the model of said test compound on the model of said active site; assessing whether said test compound model fits spatially into the active site; preparing said test compound that fits spatially into the active site; using said test compound in a biological assay for a protease characterized by said active site; and determining whether said test compound inhibits cathepsin K activity in said assay.
  • This invention involves a method of drug design comprising using the structural coordinates of a procathepsin K crystal to computationally evaluate a chemical entity for associating with the active site of cathepsin K.
  • this invention provides for a method of identifying inhibitors of cathepsin K said method comprising the steps of: providing the coordinates of the atoms of the procathepsin K active site and the coordinates of the atoms of the propeptide in the region of the active site according to Table IV to a computerized modeling system, wherein said system displays the three-dimensional structure thereof; displaying and superimposing a model of a test compound on the model of the propeptide; assessing whether said test compound interacts with the active site of procathepsin K in a manner similar to that of the propeptide; preparing said test compound that interacts with the active site of procathepsin K in a manner similar to that of the propeptide; and testing said test compound for cathepsin K inhibitory activity.
  • the three dimensional (3D) structure of the instant protease proenzyme reveals that human cathepsin K is highly homologous to other known cysteine proteinases of the papain family in the region of the mature enzyme. Cathepsin K is synthesized and secreted from the cell as an inactive proenzyme. Autoproteolytic degradation of the N-terminal 99 amino acid pro-peptide produces the active, mature form of cathepsin K. It is presumed that the activation of procathepsin K in vivo occurs in the bone resorption pit which has a low pH environment.
  • the instant invention provides for the crystal structure of human procathepsin K at 2.6 •ngstroms resolution. Knowing the structure of procathepsin K provides a basis for the design of inhibitors of cathepsin K.
  • Figure 1 Listed in Figure 1 are the known amino acid sequences for the proenzymes of the papain superfamily of cysteine proteases: cathepsin K, cathepsin S, cathepsin L, actinidin, cathepsin H and cathepsin B, aligned to illustrate the homologies there between.
  • the crystal structure of human procathepsin K has been determined at 2.6 Angstroms. The structure was determined using the method of molecular replacement and refined to an R c values of 0.204. Further refinement of the atomic coordinates will change the numbers in Table I.
  • Human procathepsin K contains 314 amino acids and the model of the enzyme provided herein is represented by residues 5 to 314. The first five amino acid residues are not well ordered in the structure.
  • the procathepsin K crystal structure reveals a conformation of the propeptide bound to the active site that is heretofor unknown and comprises a distinct three dimensional arrangement of atoms.
  • Table I discloses the protein coordinates of procathepsin K. These data are reported for the crystal structure described herein. The data are reported in Angstroms with reference to an orthogonal coordinate system in standard format, illustrating the atom, i.e., nitrogen, oxygen, carbon, sulfur (at , ⁇ , ⁇ , ⁇ , or ⁇ , positions in the amino acid residues); the amino acid residue in which the atom is located with amino acid number, and the coordinates X, Y and Z in Angstroms (A) from the crystal structure. Note that each atom in the active site and the entire structure has an unique position in the crystal. The data also report the B or Temperature Factor values, which indicate the degree of thermal motion of the atom in root mean square displacement measurements (A ⁇ ). Figure 1 illustrates the procathepsin K crystalline structure of the invention, including the active site.
  • the active site of procathepsin K and the propeptide is shown in Figure 2.
  • the pro-peptide of procathepsin K exhibits a globular domain formed by three alpha-helices and one beta-strand which is located above the right domain of cathepsin K.
  • An extended loop stretches from the globular domain through the active site and connects at the bottom of the mature cathepsin K.
  • the RMS deviation between the CA atoms of the mature cathepsin K and the CA atoms of the refined proenzyme structure is only 0.46 A (215 CA pairs).
  • Residues 74 to 80 of the pro-peptide lie in the active site and are shown in Figure 2.
  • the residues from cathepsin K which form contacts with the propeptide are shown as a ball-and-stick model.
  • the orientation of the propeptide in the active site cleft is opposite to that postulated for a natural substrate.
  • the S' subsites (located above Cys 124 in Figure 2) are occupied by Met 75 (S2') and Thr 76 (SI').
  • the S subsites are occupied by Gly 77 (SI), Leu 78 (S2) and Lys 79 (S3).
  • the main chain atoms of the propeptide form numerous hydrogen bonds to mature cathepsin K and are listed as follows: Lys 74 to Cys 121; Met 75 to Gin 118 and Trp 283; Gly 77 to Gly 165, and Lys 79 to Gly 165.
  • the sidechains of the propeptide form extensive contacts with mature cathepsin K.
  • the side chain of Met 75 is located in a hydrophobic area surrounded by side chains of Leu 63 and Met 66 from the propeptide and Trp 283 from mature cathepsin K.
  • Thr 76 forms hydrogen bonds from its sidechain hydroxyl group to mainchain carbonyls from residue Val 72 and Asn 260.
  • Thr 76 faces the hydrophobic cluster formed by residues Trp 283 and His 261.
  • Gly 77 is located at the center of the active site within van der Waals radius of the catalytic cysteine. A residue with a larger sidechain would not fit at this position.
  • Leu 78 occupies the S2 active site hydrophobic pocket, which is formed by Tyr 166, Met 167, Ala 233, Ala 262 and Leu 308. Lys 79 faces away from the catalytic site and forms a salt bridge with Asp 160.
  • the propeptide is anchored to the mature protein by hydrophobic interactions and hydrogen bonding.
  • the beta strand from the propeptide hydrogen bonds with a betastrand from mature cathepsin K forming an anti-parallel beta sheet.
  • the apolar surface of the mature enzyme is formed by residues Ser 240, Phe 241, Phe 243, Tyr 244, Tyr 249, Tyr 250, His 261, Trp 283, Trp 287 and Met 295.
  • Six of these sidechains, Phe 241, Tyr 244, Tyr 249, His 261, Trp 283, Trp 287 form an aromatic-aromatic stacking network.
  • Residues Leu 78 and Val 80 pack against residues Tyr 166, Met 167, Ala 233 and Leu 308.
  • a third region of hydrophobic interactions occurs near the C-terminus of the propeptide with residue His 84, Leu 91, Tyr 92 and He 93 packed against residue Tyr 209, He 212, Pro 231 and Phe 311 ( Figure 3).
  • Electrostatic interactions in the form of salt bridges are likely to play a significant role in the stabilization of the globular structure in the N-terminal domain of the propeptide.
  • Asp 27 and Asp65 also form important hydrogen bonds to Asn 23 and Trp 14, respectively.
  • Activation of the proenzyme occurs at low pH. Therefore disruption of these salt bridges at low pH may disrupt the globular structure and lead to auto-activation of the proenzyme.
  • the crystal structure of procathepsin K of the present invention reveals the three dimensional structure of novel active, site interactions between the atoms of the amino acid residues of the mature and propeptide listed in Table IV.
  • protease inhibitors which may be used as therapeutic agents against diseases in which inhibition of bone resorption is indicated.
  • the discovery of the procathepsin K catalytic site permits the design of potent, highly selective protease inhibitors.
  • Another aspect of this invention involves a method for identifying inhibitors of procathepsin K characterized by the crystal structure and novel active site described herein.
  • the novel protease crystal structure of the invention permits the identification of inhibitors of protease activity. Such inhibitors may bind to all or a portion of the active site of cathepsin K; or even be competitive or non-competitive inhibitors. Once identified and screened for biological activity, these inhibitors may be used therapeutically or prophylactically to block protease activity.
  • One design approach is to probe the cathepsin K of the invention with molecules composed of a variety of different chemical entities to determine optimal sites for interaction between candidate cathepsin K inhibitors and the enzyme.
  • This invention also enables the development of compounds that can isomerize to short-lived reaction intermediates in the chemical reaction of a substrate or other compound that binds to or with cathepsin K.
  • the reaction intermediates of cathepsin K can also be deduced from the reaction product in co-complex with cathepsin K.
  • Such information is useful to design improved analogues of known cysteine protease inhibitors or to design novel classes of inhibitors based on the reaction intermediates of the cathepsin K enzyme and cathepsin K inhibitor co-complex. This provides a novel route for designing cathepsin K inhibitors with both high specificity and stability.
  • cathepsin K may crystallize in more than one crystal form
  • the structure coordinates of procathepsin K, or portions thereof, as provided by this invention are particularly useful to solve the structure of those other crystal forms of cathepsin K. They may also be used to solve the structure of cathepsin K mutants, cathepsin K co-complexes, or of the crystalline form of any other protein with significant amino acid sequence homology to any functional domain of cathepsin K.
  • the unknown crystal structure whether it is another crystal form of cathepsin K, a cathepsin K mutant, or a cathepsin K co-complex, or the crystal of some other protein with significant amino acid sequence homology to any functional domain of cathepsin K, may be determined using the procathepsin K structure coordinates of this invention as provided in Table I.
  • This method will provide an accurate structural form for the unknown crystal more quickly and efficiently than attempting to determine such information ab initio.
  • the procathepsin K structure permits the screening of known molecules and/or the designing of new molecules which bind to the protease structure, particularly at the active site, via the use of computerized evaluation systems.
  • computer modelling systems are available in which the sequence of the protease, and the protease structure (i.e., atomic coordinates of procathepsin K and/or the atomic coordinate of the active site cavity, bond angles, dihedral angles, distances between atoms in the active site region, etc. as provided by Table I may be input.
  • a machine readable medium may be encoded with data representing the coordinates of Table I in this process.
  • the computer then generates structural details of the site into which a test compound should bind, thereby enabling the determination of the complementary structural details of said test compound.
  • the design of compounds that bind to or inhibit cathepsin K according to this invention generally involves consideration of two factors.
  • the compound must be capable of physically and structurally associating with cathepsin K.
  • Non-covalent molecular interactions important in the association of cathepsin K with its substrate include hydrogen bonding, van der Waals and hydrophobic interactions.
  • the compound must be able to assume a conformation that allows it to associate with cathepsin K. Although certain portions of the compound will not directly participate in this association with cathepsin K, those portions may still influence the overall conformation of the molecule. This, in turn, may have a significant impact on potency.
  • Such conf ⁇ rmational requirements include the overall three-dimensional structure and orientation of the chemical entity or compound in relation to all or a portion of the binding site, e.g., active site or accessory binding site of cathepsin K, or the spacing between functional groups of a compound comprising several chemical entities that directly interact with cathepsin K.
  • the potential inhibitory or binding effect of a chemical compound with cathepsin K may be estimated prior to its actual synthesis and testing by the use of computer modelling techniques. If the theoretical structure of the given compound suggests insufficient interaction and association between it and cathepsin K, synthesis and testing of the compound is obviated. However, if computer modelling indicates a strong interaction, the molecule may then be synthesized and tested for its ability to bind to cathepsin K in a suitable assay. In this manner, synthesis of inoperative compounds may be avoided.
  • An inhibitory or other binding compound of cathepsin K may be computationally evaluated and designed by means of a series of steps in which chemical entities or fragments are screened and selected for their ability to associate with the individual binding pockets or other areas of cathepsin K.
  • One skilled in the art may use one of several methods to screen chemical entities or fragments for their ability to associate with cathepsin K and more particularly with the individual binding pockets of the cathepsin K active site or accessory binding site.
  • This process may begin by visual inspection of, for example, the active site on the computer screen based on the cathepsin K coordinates in Table I. Selected fragments or chemical entities may then be position cathepsin K. Docking may be accomplished using software such as Quanta and Sybyl, followed by energy minimization and molecular dynamics with standard molecular mechanics forcefields, such as CHARMM and AMBER.
  • Specialized computer programs may also assist in the process of selecting fragments or chemical entities. These include: • GRID [P. J. Goodford, "A Computational Procedure for Determining Energetically Favorable Binding Sites on Biologically Important Macromolecules", J. Med. Chem.. 28:849-857 (1985)]. GRID is available from Oxford University, Oxford, UK. • MCSS [A. Miranker and M. Karplus, "Functionality Maps of Binding Sites: A Multiple Copy Simultaneous Search Method", Proteins: Structure. Function and Genetics.11:29-34 (1991)]. MCSS is available from Molecular Simulations, Burlington, MA. • AUTODOCK [D. S. Goodsell and A. J.
  • DOCK [I. D. Kuntz et al, "A Geometric Approach to Macromolecule- Ligand Interactions". J. Mol. Biol.. 161:269-288 (1982)1. DOCK is available from
  • CAVEAT A Program to Facilitate the Structure-Derived Design of Biologically Active Molecules", in Molecular Recognition in Chemical and Biological Problems". Speical Pub., Royal Chem. Soc. 78, pp. 182-196 (1989)]. CAVEAT is available from the University of California, Berkeley, CA.
  • 3D Database systems such as MACCS-3D (MDL Information Systems, San Leandro, CA). This area is reviewed in Y. C. Martin, "3D Database Searching in Drug Design", J. Med. Chem.. 35:2145-2154 (1992).
  • inhibitory or other type of binding compounds may be designed as a whole or "de novo" using either an empty active site or optionally including some portion(s) of a known inhibitor(s).
  • LUDI H.-J. Bohm, "The Computer Program LUDI: A New Method for the De Novo Design of Enzyme Inhibitors", J. Comp. Aid. Molec. Design. 6:61- 78 (1992)].
  • LUDI is available from Biosym Technologies, San Diego, CA.
  • LEGEND [Y. Nishibata and A. Itai, Tetrahedron. 47:8985 (1991)]. LEGEND is available from Molecular Simulations, Burlington, MA.
  • model building techniques and computer evaluation systems described herein are not a limitation on the present invention.
  • a large number of compounds may be quickly and easily examined and expensive and lengthy biochemical testing avoided.
  • the need for actual synthesis of many compounds is effectively eliminated.
  • the protease inhibitor may be tested for bioactivity using standard techniques.
  • structure of the invention may be used in binding assays using conventional formats to screen inhibitors.
  • Suitable assays for use herein include, but are not limited to, the enzyme- linked immunosorbent assay (ELISA), or a fluoresence quench assay. See, for example, the cathepsin K activity assay of Example 2 below.
  • ELISA enzyme- linked immunosorbent assay
  • fluoresence quench assay See, for example, the cathepsin K activity assay of Example 2 below.
  • Other assay formats may be used; these assay formats are not a limitation on the present invention.
  • the protease structure of the invention permit the design and identification of synthetic compounds and/or other molecules which have a shape complimentary to the conformation of the protease active site of the invention.
  • the coordinates of the protease structure of the invention may be provided in machine readable form, the test compounds designed and/or screened and their conformations superimposed on the structure of the protease of the invention. Subsequently, suitable candidates identified as above may be screened for the desired protease inhibitory bioactivity, stability, and the like.
  • inhibitors may be used therapeutically or prophylactically to block cathepsin K activity.
  • Procathepsin K (procat K) was expressed using the PET system (Novagen).
  • Procathepsin K an existing clone containing the full-length human preprocathepsinK cDNA was subject to PCR amplifcation using oligos designed to introduce a Met residue at amino acid 18 of the full-length preproenzyme. The resulting fragment was cleaved with Ncol and
  • the washed pellet was resuspended into 150 mL of the same buffer prepared at 4 C and lysed by passage twice through a cell disruptor (Avestin) at 10,000 psi.
  • the lysate was centrifuged as above, the supernatant decanted and the pellet suspended in 150 mL 50 mM Tris/HCl, 6 M guanidine HC1, 10 mM DTT, 5 mM EDTA, 150 mM NaCl, pH 8.0.
  • insoluble cellular debris was removed by centrifugation at 23,000 x g for 30 mins and the supernatant clarified by filtration
  • Crystals of procathepsin K grew to a size of approximately 0.3 mm ⁇ in ten days at 4° C.
  • the concentration of procathepsin K used in the crystallization was lOmg/ml.
  • the hanging drops vapour diffusion method was used with a solution of 8-10% PEG6000, 0.1M Na citrate at pH 5.0.
  • the crystals contain one molecule in the asymmetric unit and contain 60% solvent.
  • procathepsin K was solved by molecular replacement using the program XPLOR (Brunger, A. T., Kuriyan, J. and Karplus, M. (1987) Science 235, 458-460).
  • the coordinates for 217 residues from the structure of human cathespin K (Zhao, B. et al. (1977) Nature Struct. Biol. 4, 109-111) were used as a starting model.
  • the search was carried out with pseudo-orthogonal euler angles in Patterson space from ⁇ l 0-180°,
  • the highest peak was 15.7 ⁇ .
  • the translation search was computed using data from 10 -3.5 A and gave a peak at 26 ⁇ .
  • the resulting model yielded a R-factor of 0.43.
  • the R-factor for the model dropped to 0.40 after rigid body refinement from 10-3.5 A and to 0.31 after 200 cycles of positional refinement.
  • the electron density maps were improved using solvent flattening and by histogram matching as implemented in the module DM of the CCP4 suite of programs (Collaborative Computational Project, Number 4 (1994), "The CCP4 Suite: Programs for Protein Crystallography", Acta Cryst. D50, 760-763).
  • the modified electron density map was to build the model of used procathepsin K.
  • Conventional positional refinement was used to refine the structure during model building.
  • the structure was refined using X-plor.
  • the final model of procathepsin K consists of residues 5 to 214.
  • the simulated annealing refinement was finally carried out to a final R of 0.204.
  • the backbone conformation of all residues is within the Ramachandran plot limits.
  • v is the velocity of the reaction with maximal velocity V m
  • A is the concentration of substrate with Michaelis constant of K a
  • I is the concentration of inhibitor.
  • [AMC] v S s t + (vo - v ss ) [1 - exp (-kobst)J ' 'k ⁇ bs (2)
  • This assay measures the affinity of inhibitors to cathepsin K.
  • One skilled in the art would consider any compound exhibiting a K j value of less than 50 micromolar to be a potential lead compound for further research.
  • the cells were washed x2 with cold RPMI-1640 by centrifugation (1000 rpm, 5 min at 4°C) and then transferred to a sterile 15 mL centrifuge tube. The number of mononuclear cells were enumerated in an improved Neubauer counting chamber.
  • Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG, were removed from their stock bottle and placed into 5 mL of fresh medium (this washes away the toxic azide preservative). The medium was removed by immobilizing the beads on a magnet and is replaced with fresh medium. The beads were mixed with the cells and the suspension was incubated for 30 min on ice. The suspension was mixed frequently. The bead-coated cells were immobilized on a magnet and the remaining cells (osteoclast-rich fraction) were decanted into a sterile 50 mL centrifuge tube. Fresh medium was added to the bead- coated cells to dislodge any trapped osteoclasts. This wash process was repeated lO. The bead-coated cells were discarded.
  • the osteoclasts were enumerated in a counting chamber, using a large-bore disposable plastic Pasteur pipette to charge the chamber with the sample.
  • the cells were pelleted by centrifugation and the density of osteoclasts adjusted to 1.5xl ⁇ 4/mL in EMEM medium, supplemented with 10% fetal calf serum and 1.7g/liter of sodium bicarbonate. 3 mL aliquots of the cell suspension ( per treatment) were decanted into 15 mL centrifuge tubes. These cells were pelleted by centrifugation. To each tube 3 mL of the appropriate treatment was added (diluted to 50 uM in the EMEM medium).
  • a positive control (87MEM1 diluted to 100 ug/mL) and an isotype control (IgG2a diluted to 100 ug/mL).
  • the tubes were incubate at 37°C for 30 min.
  • 0.5 mL aliquots of the cells were seeded onto sterile dentine slices in a 48- well plate and incubated at 37°C for 2 h. Each treatment was screened in quadruplicate.
  • the slices were washed in six changes of warm PBS (10 mL / well in a 6-well plate) and then placed into fresh treatment or control and incubated at 37°C for 48 h.
  • the slices were then washed in phosphate buffered saline and fixed in 2% glutaraldehyde (in 0.2M sodium cacodylate) for 5 min., following which they were washed in water and incubated in buffer for 5 min at 37°C.
  • the slices were then washed in cold water and incubated in cold acetate buffer / fast red garnet for 5 min at 4°C. Excess buffer was aspirated, and the slices were air dried following a wash in water.
  • the TRAP positive osteoclasts were enumerated by bright-field microscopy and were then removed from the surface of the dentine by sonication. Pit volumes were determined using the Nikon/Lasertec ILM21W confocal microscope.
  • the three dimensional atomic structure can be readily used as a template for selecting potent inhibitors.
  • Various computer programs and databases are available for the purpose.
  • a good inhibitor should at least have excellent steric and electrostatic complementarity to the target, a fair amount of hydrophobic surface buried and sufficent conformational rigidity to minimize entropy loss upon binding.
  • the approach usually comprises several steps: 1) Define a region to target, the active site cavity of procathepsin K can be selected, but any place that is essential to the protease activity could become a potential target. Since the crystal structure has been determined, the spatial and chemical properties of the target region is known.
  • a limited numer of promising compounds can be selected through the process. They can then be synthesized and assayed for their inhibitory properties. The success rate can sometimes be as high as 20%, and it may still be higher with the rapid progresses in computing methods.
  • R C ⁇ I(F 0 - F C )I / F 0
  • F c calculated structure amplitude EDTA, ethylenediaminetetraacetic acid DTT, 1,4-dithiothreitol SDS-PAGE, sodium dodecylsulfate polyacrylamide gel electrophoresis
  • Table I Table of the orthogonal three dimensional coordinates in Angstroms and B factors (A 2 ) for procathepsin K.

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Abstract

A novel procathepsin K crystalline structure is identified. Also disclosed are methods of identifying inhibitors of cathepsin K and methods of designing inhibitors of cathepsin K using structural and physical data based on the crystallographic structure of procathepsin K.

Description

METHOD OF INHIBITING CATHEPSIN K
Field of the Invention This invention relates to a method of inhibiting cathepsin K by the utilization of the physical and spacial characteristics that allow for the interaction of the propeptide with the active site of the enzyme. This interaction inhibits the activity of cathepsin K and this information is useful in the design of specific chemical compounds for treating diseases in which said inhibition is indicated, such as osteoporosis and periodontal disease. This invention also relates to a novel crystalline structure of procathepsin K, the identification of the propeptide bound to the catalytic active site for this enzyme and methods enabling the design and selection of inhibitors of said active site.
Background of the Invention
Cathepsin K is a member of the family of enzymes which are part of the papain superfamily of cysteine proteases. Cathepsins B, H, L, N and S have been described in the literature. Recently, cathepsin K polypeptide and the cDNA encoding such polypeptide were disclosed in U.S. Patent No. 5,501,969 (called cathepsin O therein). Cathepsin K has been recently expressed, purified, and characterized. Bossard, M. J., et al., (1996) J. Biol. Chem. 271, 12517-12524; Drake, F.H., et al., (1996) J. Biol. Chem. 271, 12511-12516; Bromme, D., et al., (1996) J. Biol. Chem. Ill, 2126-2132.
Cathepsin K has been variously denoted as cathepsin O, cathepsin X or cathepsin O2 in the literature. The designation cathepsin K is considered to be the more appropriate one (name assigned by Nomenclature Committee of the International Union of Biochemistry and Molecular Biology).
Cathepsins of the papain superfamily of cysteine proteases function in the normal physiological process of protein degradation in animals, including humans, e.g., in the degradation of connective tissue. However, elevated levels of these enzymes in the body can result in pathological conditions leading to disease. Thus, cathepsins have been implicated in various disease states, including but not limited to, infections by pneumocystis carinii, trypsanoma cruzi, trypsanoma brucei brucei, and Crithidia fusiculata; as well as in schistosomiasis malaria, tumor metastasis, metachromatic leukodystrophy, muscular dystrophy, amytrophy, and the like. See International Publication Number WO 94/04172, published on March 3, 1994, and references cited therein. See also European Patent Application EP 0 603 873 Al, and references cited therein. Two bacterial cysteine proteases from P. gingivallis, called gingipains, have been implicated in the pathogenesis of gingivitis. Potempa, J., et al. (1994) Perspectives in Drug Discovery and Design, 2, 445-458.
Cathepsin K is believed to play a causative role in diseases of excessive bone or cartilage loss. Bone is composed of a protein matrix in which spindle- or plate- shaped crystals of hydroxyapatite are incorporated. Type I Collagen represents the major structural protein of bone comprising approximately 90% of the structural protein. The remaining 10% of matrix is composed of a number of non-collagenous proteins, including osteocalcin, proteoglycans, osteopontin, osteonectin, thrombospondin, fibronectin, and bone sialoprotein. Skeletal bone undergoes remodeling at discrete foci throughout life. These foci, or remodeling units, undergo a cycle consisting of a bone resorption phase followed by a phase of bone replacement.
Bone resorption is carried out by osteoclasts, which are multinuclear cells of hematopoietic lineage. The osteoclasts adhere to the bone surface and form a tight sealing zone, followed by extensive membrane ruffling on their apical (i.e., resorbing) surface. This creates an enclosed extracellular compartment on the bone surface that is acidified by proton pumps in the ruffled membrane, and into which the osteoclast secretes proteolytic enzymes. The low pH of the compartment dissolves hydroxyapatite crystals at the bone surface, while the proteolytic enzymes digest the protein matrix. In this way, a resorption lacuna, or pit, is formed. At the end of this phase of the cycle, osteoblasts lay down a new protein matrix that is subsequently mineralized. In several disease states, such as osteoporosis and Paget's disease, the normal balance between bone resorption and formation is disrupted, and there is a net loss of bone at each cycle. Ultimately, this leads to weakening of the bone and may result in increased fracture risk with minimal trauma.
The abundant selective expression of cathepsin K in osteoclasts strongly suggests that this enzyme is essential for bone resorption. Thus, selective inhibition of cathepsin K may provide an effective treatment for diseases of excessive bone loss, including, but not limited to, osteoporosis, gingival diseases such as gingivitis and periodontitis, Paget's disease, hypercalcemia of malignancy, and metabolic bone disease. Cathepsin K levels have also been demonstrated to be elevated in chondroclasts of osteoarthritic synovium. Thus, selective inhibition of cathepsin K may also be useful for treating diseases of excessive cartilage or matrix degradation, including, but not limited to, osteoarthritis and rheumatoid arthritis. Metastatic neoplastic cells also typically express high levels of proteolytic enzymes that degrade the surrounding matrix. Thus, selective inhibition of cathepsin K may also be useful for treating certain neoplastic diseases.
Surprisingly, the propeptide of cathepsin K exhibits characteristics that allow for the interaction of the propeptide with specific residues of the active site of cathepsin K and this information is useful in the design of specific chemical compounds as inhibitors of cathepsin K and would be useful for treating diseases in which inhibition of bone resorption is indicated, such as osteoporosis and periodontal disease. Thus, this invention relates to the method of inhibiting cathepsin K using the crystal structure of procathepsin K hereinbelow defined.
Summary of the Invention In one aspect, the present invention provides a method for designing compounds to inhibit cathepsin K with certain structural, physical and spacial characteristics that allow for the interaction of said compounds with specific residues of the active site of the enzyme in a manner similar to the interaction of the propeptide with the enzyme active site. This interaction inhibits the activity of cathepsin K and, thus, treats diseases in which bone resorption is a factor.
In another aspect, the present invention provides a novel cysteine protease proenzyme in crystalline form, in particular this invention relates to a novel procathepsin K crystal.
In yet another aspect, the invention provides a novel protease proenzyme composition, specifically a novel procathepsin K composition characterized by a three dimensional catalytic site inhibited by the atoms of the propeptide residues listed in Table IV. In still another aspect, the invention provides a method for identifying inhibitors said method comprising the steps of: providing the coordinates of the proenzyme structure of the invention to a computerized modeling system; and identifying compounds which will bind to the structure in a manner similar to that of the propeptide. Other aspects and advantages of the present invention are described further in the following detailed description of the preferred embodiments thereof.
Brief Description of the Drawings Figure 1 is the amino acid sequence of procathepsin K aligned with the amino acid sequences of other cysteine protease proenzymes. Figure 2 is a ribbon diagram of procathepsin K. The amino and carboxyl- termini are indicated by N and C. The drawing was produced using the program MOLSCRIPT [Kraulis, P., J. Appl. Crystallogr., 24, 946-950 (1991)].
Figure 3 is an illustration of the active site of procathepsin K. Table I provides the three dimensional protein coordinates of the procathepsin K crystalline structure of the invention.
Table II provides a listing of the three atom angles between atoms of the propeptide and the protein for all propeptide atoms within 5 Angstroms of the protein. Table III provides a listing of the distances between atoms of the propeptide and the protein for all propeptide atoms within 5 Angstroms of the protein.
Table IV provides the atoms of the amino acid residues of the catalytic site and the propeptide amino acid residues which interact with them.
Detailed Description of the Invention
The present invention provides a novel cysteine protease proenzyme crystalline structure, and methods of use of the crystalline form and active site interactions with the inhibitory propeptide to identify protease inhibitor compounds. In particular, the present invention provides a method for inhibiting cathepsin K by designing compounds with certain structural, physical and spacial characteristics that allow for the interaction of said compounds with specific residues of the active site of the enzyme based on the structure of the proenzyme. This interaction inhibits the activity of cathepsin K and, thus, treats diseases in which bone resorption is a factor. In another aspect, the present invention provides a novel cysteine protease proenzyme in crystalline form, in particular procathepsin K in crystalline form as defined by the positions in Table I herein.
In still another aspect, the present invention provides a novel protease proenzyme composition, specifically a novel procathepsin K composition, characterized by a three dimensional catalytic site formed by the atoms of the amino acid residues listed in Table IV herein.
A key aspect of this invention provides a method of inhibiting cathepsin K which comprises administering to a mammal in need thereof a compound that mimics the interaction of the propeptide with the residues of the active site of procathepsin K. Specifially, this compound comprises: (i) an oxygen atom which interacts with a hydrogen atom donated by the indole nitrogen of tryptophan 283 wherein the distance between these two atoms is 2.7-3.5A;
(ii) a hydrophobic group that interacts with tryptophan 283 wherein the distance between the centroid of said hydrophobic group and the centroid of the side chain atoms of tryptophan 283 is 4.10-7.10A;
(iii) a hydrophobic group that interacts with tyrosine 166 , methionine 167, alanine 233, leucine 259, and leucine 308, creating a hydrophobic pocket, and has distance ranges between the centroid of said hydrophobic group and the centroids of the side chain atoms of the amino acid residues of said hydrophobic pocket which are tyrosine 166: 4.91- 5.91A, methionine 167: 5.74-6.74A, alanine 233: 4.15-5.15A, leucine 259: 6.18-7.18A, and leucine 308: 5.71-6.7lA; and
(iv) an amino group with a pKa of less than 7 or an oxygen atom, each of which interacts with a hydrogen atom donated by the amide nitrogen of glycine 264 wherein the distance between these two atoms is 2.7-3.5 A. In particular, the preferred compound consists of a hydrophobic group that interacts with said hydrophobic pocket wherein said hydrophobic group is an isobutyl group.
The method of the instant invention comprises compounds characterized by the interactions of the atoms of the procathepsin K active site and the atoms of the propeptide in the region of the acive site according to Table IV.
This invention also includes a composition comprising procathepsin K in crystalline form wherein the procathepsin K in crystalline form is characterized by structure coordinates according to Table I. In particular, the composition has an active site cavity formed by the amino acids in Table IV wherein said active site is characterized by the distance between the atoms of Table II and the bond angles between interresidue atoms of Table III.
In another aspect of this invention is an isolated, properly folded procathepsin K molecule or fragment thereof having a conformation comprising a catalytically active site formed by the residues listed in Table IV, said active site defined by the protein coordinates of Table I and a peptide, peptidomimetic or synthetic molecule which binds with the active site cavity of procathepsin K.
Importantly, this invention provides for a method of identifying an inhibitor compound capable of binding to, and inhibiting the proteolytic activity of, cathepsin K, said method comprising: introducing into a suitable computer program information defining an active site conformation of a procathepsin K molecule comprising a catalytically active site formed by the residues listed in Table IV, said active site defined by the protein coordinates of Table I, wherein said program displays the three-dimensional structure thereof; creating a three dimensional representation of the active site cavity in said computer program; displaying and superimposing the model of said test compound on the model of said active site; assessing whether said test compound model fits spatially into the active site; preparing said test compound that fits spatially into the active site; using said test compound in a biological assay for a protease characterized by said active site; and determining whether said test compound inhibits cathepsin K activity in said assay.
This invention involves a method of drug design comprising using the structural coordinates of a procathepsin K crystal to computationally evaluate a chemical entity for associating with the active site of cathepsin K.
Alternately, this invention provides for a method of identifying inhibitors of cathepsin K said method comprising the steps of: providing the coordinates of the atoms of the procathepsin K active site and the coordinates of the atoms of the propeptide in the region of the active site according to Table IV to a computerized modeling system, wherein said system displays the three-dimensional structure thereof; displaying and superimposing a model of a test compound on the model of the propeptide; assessing whether said test compound interacts with the active site of procathepsin K in a manner similar to that of the propeptide; preparing said test compound that interacts with the active site of procathepsin K in a manner similar to that of the propeptide; and testing said test compound for cathepsin K inhibitory activity. The three dimensional (3D) structure of the instant protease proenzyme reveals that human cathepsin K is highly homologous to other known cysteine proteinases of the papain family in the region of the mature enzyme. Cathepsin K is synthesized and secreted from the cell as an inactive proenzyme. Autoproteolytic degradation of the N-terminal 99 amino acid pro-peptide produces the active, mature form of cathepsin K. It is presumed that the activation of procathepsin K in vivo occurs in the bone resorption pit which has a low pH environment. The instant invention provides for the crystal structure of human procathepsin K at 2.6 •ngstroms resolution. Knowing the structure of procathepsin K provides a basis for the design of inhibitors of cathepsin K.
Listed in Figure 1 are the known amino acid sequences for the proenzymes of the papain superfamily of cysteine proteases: cathepsin K, cathepsin S, cathepsin L, actinidin, cathepsin H and cathepsin B, aligned to illustrate the homologies there between. According to the present invention the crystal structure of human procathepsin K has been determined at 2.6 Angstroms. The structure was determined using the method of molecular replacement and refined to an Rc values of 0.204. Further refinement of the atomic coordinates will change the numbers in Table I. Refinement of the crystal structure from another crystal form will result in a new set of coordinates, determination of the crystal structure of another cysteine protease will also result in different set of numbers for coordinates in Table I which has an experimental error of approximately 0.4 Angstroms. Also, for example, the amino acid sequence of the cysteine proteases can be varied by mutation derivatization or by use of a different source of the protein.
Human procathepsin K contains 314 amino acids and the model of the enzyme provided herein is represented by residues 5 to 314. The first five amino acid residues are not well ordered in the structure.
The procathepsin K crystal structure reveals a conformation of the propeptide bound to the active site that is heretofor unknown and comprises a distinct three dimensional arrangement of atoms.
Table I discloses the protein coordinates of procathepsin K. These data are reported for the crystal structure described herein. The data are reported in Angstroms with reference to an orthogonal coordinate system in standard format, illustrating the atom, i.e., nitrogen, oxygen, carbon, sulfur (at , β, γ, δ, or ε, positions in the amino acid residues); the amino acid residue in which the atom is located with amino acid number, and the coordinates X, Y and Z in Angstroms (A) from the crystal structure. Note that each atom in the active site and the entire structure has an unique position in the crystal. The data also report the B or Temperature Factor values, which indicate the degree of thermal motion of the atom in root mean square displacement measurements (A^). Figure 1 illustrates the procathepsin K crystalline structure of the invention, including the active site.
The active site of procathepsin K and the propeptide is shown in Figure 2. The pro-peptide of procathepsin K exhibits a globular domain formed by three alpha-helices and one beta-strand which is located above the right domain of cathepsin K. An extended loop stretches from the globular domain through the active site and connects at the bottom of the mature cathepsin K. The RMS deviation between the CA atoms of the mature cathepsin K and the CA atoms of the refined proenzyme structure is only 0.46 A (215 CA pairs).
Residues 74 to 80 of the pro-peptide lie in the active site and are shown in Figure 2. The residues from cathepsin K which form contacts with the propeptide are shown as a ball-and-stick model. The orientation of the propeptide in the active site cleft is opposite to that postulated for a natural substrate. The S' subsites (located above Cys 124 in Figure 2) are occupied by Met 75 (S2') and Thr 76 (SI'). The S subsites are occupied by Gly 77 (SI), Leu 78 (S2) and Lys 79 (S3). The main chain atoms of the propeptide form numerous hydrogen bonds to mature cathepsin K and are listed as follows: Lys 74 to Cys 121; Met 75 to Gin 118 and Trp 283; Gly 77 to Gly 165, and Lys 79 to Gly 165. The sidechains of the propeptide form extensive contacts with mature cathepsin K. The side chain of Met 75 is located in a hydrophobic area surrounded by side chains of Leu 63 and Met 66 from the propeptide and Trp 283 from mature cathepsin K. Thr 76 forms hydrogen bonds from its sidechain hydroxyl group to mainchain carbonyls from residue Val 72 and Asn 260. Additionally, the side-chain methyl group of Thr 76 faces the hydrophobic cluster formed by residues Trp 283 and His 261. Gly 77 is located at the center of the active site within van der Waals radius of the catalytic cysteine. A residue with a larger sidechain would not fit at this position. Leu 78 occupies the S2 active site hydrophobic pocket, which is formed by Tyr 166, Met 167, Ala 233, Ala 262 and Leu 308. Lys 79 faces away from the catalytic site and forms a salt bridge with Asp 160.
The propeptide is anchored to the mature protein by hydrophobic interactions and hydrogen bonding. The hydrophobic face of helices 2 and 3, and a beta strand pack against the apolar surface of mature cathepsin K (Figure 3). The beta strand from the propeptide hydrogen bonds with a betastrand from mature cathepsin K forming an anti-parallel beta sheet. The following residues from helices 2 and 3 and the beta-strand; Tyr 41, He 42, His 45, Ala 49, His 54, Tyr 56, Leu 58, Met 60, His 62, Leu 63, Met 66, Val 71, Val 72 and Met 75, form hydrophobic contacts with side chain atoms od mature cathepsin K. The apolar surface of the mature enzyme is formed by residues Ser 240, Phe 241, Phe 243, Tyr 244, Tyr 249, Tyr 250, His 261, Trp 283, Trp 287 and Met 295. Six of these sidechains, Phe 241, Tyr 244, Tyr 249, His 261, Trp 283, Trp 287 form an aromatic-aromatic stacking network. Near the active site a second region of apolar interactions exists. Residues Leu 78 and Val 80 pack against residues Tyr 166, Met 167, Ala 233 and Leu 308. A third region of hydrophobic interactions occurs near the C-terminus of the propeptide with residue His 84, Leu 91, Tyr 92 and He 93 packed against residue Tyr 209, He 212, Pro 231 and Phe 311 (Figure 3).
Electrostatic interactions in the form of salt bridges are likely to play a significant role in the stabilization of the globular structure in the N-terminal domain of the propeptide. There are seven salt bridges within the pro-peptide. These are Asp 8 to Arg 32, Lys 20 to Asp 65, Asn 24 to Asp 27, Lys 25 to Glu 28, Arg 31 to Glu 70, Arg 32 to Glu 36 and Glu 48 to His 54. Asp 27 and Asp65 also form important hydrogen bonds to Asn 23 and Trp 14, respectively. Activation of the proenzyme occurs at low pH. Therefore disruption of these salt bridges at low pH may disrupt the globular structure and lead to auto-activation of the proenzyme.
The crystal structure of procathepsin K of the present invention reveals the three dimensional structure of novel active, site interactions between the atoms of the amino acid residues of the mature and propeptide listed in Table IV.
This structure is clearly useful in the structure-based design of protease inhibitors, which may be used as therapeutic agents against diseases in which inhibition of bone resorption is indicated. The discovery of the procathepsin K catalytic site permits the design of potent, highly selective protease inhibitors.
Another aspect of this invention involves a method for identifying inhibitors of procathepsin K characterized by the crystal structure and novel active site described herein. The novel protease crystal structure of the invention permits the identification of inhibitors of protease activity. Such inhibitors may bind to all or a portion of the active site of cathepsin K; or even be competitive or non-competitive inhibitors. Once identified and screened for biological activity, these inhibitors may be used therapeutically or prophylactically to block protease activity. One design approach is to probe the cathepsin K of the invention with molecules composed of a variety of different chemical entities to determine optimal sites for interaction between candidate cathepsin K inhibitors and the enzyme. For example, high resolution X-ray diffraction data collected from crystals saturated with solvent allows the determination of where each type of solvent molecule sticks. Small molecules that bind tightly to those sites can then be designed and synthesized and tested for their cathepsin K inhibitor activity.
This invention also enables the development of compounds that can isomerize to short-lived reaction intermediates in the chemical reaction of a substrate or other compound that binds to or with cathepsin K. Thus, the time-dependent analysis of structural changes in cathepsin K during its interaction with other molecules is permitted. The reaction intermediates of cathepsin K can also be deduced from the reaction product in co-complex with cathepsin K. Such information is useful to design improved analogues of known cysteine protease inhibitors or to design novel classes of inhibitors based on the reaction intermediates of the cathepsin K enzyme and cathepsin K inhibitor co-complex. This provides a novel route for designing cathepsin K inhibitors with both high specificity and stability.
Because cathepsin K may crystallize in more than one crystal form, the structure coordinates of procathepsin K, or portions thereof, as provided by this invention are particularly useful to solve the structure of those other crystal forms of cathepsin K. They may also be used to solve the structure of cathepsin K mutants, cathepsin K co-complexes, or of the crystalline form of any other protein with significant amino acid sequence homology to any functional domain of cathepsin K.
One method that may be employed for this purpose is molecular replacement. In this method, the unknown crystal structure, whether it is another crystal form of cathepsin K, a cathepsin K mutant, or a cathepsin K co-complex, or the crystal of some other protein with significant amino acid sequence homology to any functional domain of cathepsin K, may be determined using the procathepsin K structure coordinates of this invention as provided in Table I. This method will provide an accurate structural form for the unknown crystal more quickly and efficiently than attempting to determine such information ab initio. Thus, the procathepsin K structure provided herein permits the screening of known molecules and/or the designing of new molecules which bind to the protease structure, particularly at the active site, via the use of computerized evaluation systems. For example, computer modelling systems are available in which the sequence of the protease, and the protease structure (i.e., atomic coordinates of procathepsin K and/or the atomic coordinate of the active site cavity, bond angles, dihedral angles, distances between atoms in the active site region, etc. as provided by Table I may be input. Thus, a machine readable medium may be encoded with data representing the coordinates of Table I in this process. The computer then generates structural details of the site into which a test compound should bind, thereby enabling the determination of the complementary structural details of said test compound.
More particularly, the design of compounds that bind to or inhibit cathepsin K according to this invention generally involves consideration of two factors. First, the compound must be capable of physically and structurally associating with cathepsin K. Non-covalent molecular interactions important in the association of cathepsin K with its substrate include hydrogen bonding, van der Waals and hydrophobic interactions. Second, the compound must be able to assume a conformation that allows it to associate with cathepsin K. Although certain portions of the compound will not directly participate in this association with cathepsin K, those portions may still influence the overall conformation of the molecule. This, in turn, may have a significant impact on potency. Such confαrmational requirements include the overall three-dimensional structure and orientation of the chemical entity or compound in relation to all or a portion of the binding site, e.g., active site or accessory binding site of cathepsin K, or the spacing between functional groups of a compound comprising several chemical entities that directly interact with cathepsin K.
The potential inhibitory or binding effect of a chemical compound with cathepsin K may be estimated prior to its actual synthesis and testing by the use of computer modelling techniques. If the theoretical structure of the given compound suggests insufficient interaction and association between it and cathepsin K, synthesis and testing of the compound is obviated. However, if computer modelling indicates a strong interaction, the molecule may then be synthesized and tested for its ability to bind to cathepsin K in a suitable assay. In this manner, synthesis of inoperative compounds may be avoided.
An inhibitory or other binding compound of cathepsin K may be computationally evaluated and designed by means of a series of steps in which chemical entities or fragments are screened and selected for their ability to associate with the individual binding pockets or other areas of cathepsin K.
One skilled in the art may use one of several methods to screen chemical entities or fragments for their ability to associate with cathepsin K and more particularly with the individual binding pockets of the cathepsin K active site or accessory binding site. This process may begin by visual inspection of, for example, the active site on the computer screen based on the cathepsin K coordinates in Table I. Selected fragments or chemical entities may then be position cathepsin K. Docking may be accomplished using software such as Quanta and Sybyl, followed by energy minimization and molecular dynamics with standard molecular mechanics forcefields, such as CHARMM and AMBER.
Specialized computer programs may also assist in the process of selecting fragments or chemical entities. These include: • GRID [P. J. Goodford, "A Computational Procedure for Determining Energetically Favorable Binding Sites on Biologically Important Macromolecules", J. Med. Chem.. 28:849-857 (1985)]. GRID is available from Oxford University, Oxford, UK. • MCSS [A. Miranker and M. Karplus, "Functionality Maps of Binding Sites: A Multiple Copy Simultaneous Search Method", Proteins: Structure. Function and Genetics.11:29-34 (1991)]. MCSS is available from Molecular Simulations, Burlington, MA. • AUTODOCK [D. S. Goodsell and A. J. Olsen, "Automated Docking of Substrates to Proteins by Simulated Annealing", Proteins: Structure. Function, and Genetics. 8:195-202 (1990)]. AUTODOCK is available from Scripps Research Institute, La Jolla, CA.
• DOCK [I. D. Kuntz et al, "A Geometric Approach to Macromolecule- Ligand Interactions". J. Mol. Biol.. 161:269-288 (1982)1. DOCK is available from
University of California, San Francisco, CA.
Additional commercially available computer databases for small molecular compounds includes Cambridge Strucutral Database and Fine Chemical Database, for a review see Rusinko, A., Chem. Des. Auto. News 8, 44-47 (1993). Once suitable chemical entities or fragments have been selected, they can be assembled into a single compound or inhibitor. Assembly may be proceeded by visual inspection of the relationship of the fragments to each other on the three- dimensional image displayed on a computer screen in relation to the structure coordinates of cathepsin K. This would be followed by manual model building using software such as Quanta or Sybyl.
Useful programs to aid one of skill in the art in connecting the individual chemical entities or fragments include:
• CAVEAT [P. A. Bartlett et al, "CAVEAT: A Program to Facilitate the Structure-Derived Design of Biologically Active Molecules", in Molecular Recognition in Chemical and Biological Problems". Speical Pub., Royal Chem. Soc. 78, pp. 182-196 (1989)]. CAVEAT is available from the University of California, Berkeley, CA.
• 3D Database systems such as MACCS-3D (MDL Information Systems, San Leandro, CA). This area is reviewed in Y. C. Martin, "3D Database Searching in Drug Design", J. Med. Chem.. 35:2145-2154 (1992).
• HOOK (available from Molecular Simulations, Burlington, MA). Instead of proceeding to build a cathepsin K inhibitor in a step-wise fashion one fragment or chemical entity at a time as described above, inhibitory or other type of binding compounds may be designed as a whole or "de novo" using either an empty active site or optionally including some portion(s) of a known inhibitor(s). These methods include: • LUDI [H.-J. Bohm, "The Computer Program LUDI: A New Method for the De Novo Design of Enzyme Inhibitors", J. Comp. Aid. Molec. Design. 6:61- 78 (1992)]. LUDI is available from Biosym Technologies, San Diego, CA.
• LEGEND [Y. Nishibata and A. Itai, Tetrahedron. 47:8985 (1991)]. LEGEND is available from Molecular Simulations, Burlington, MA.
• LeapFrog (available from Tripos Associates, St. Louis, MO). Other molecular modelling techniques may also be employed in accordance with this invention. See, e.g., N. C. Cohen et al, "Molecular Modeling Software and Methods for Medicinal Chemistry", J. Med. Chem.. 33:883-894 (1990). See also, M. A. Navia and M. A. Murcko, "The Use of Structural Information in Drug
Design", Current Opinions in Structural Biology. 2:202-210 (1992). For example, where the structures of test compounds are known, a model of the test compound may be superimposed over the model of the structure of the invention. Numerous methods and techniques are known in the art for performing this step, any of which may be used. See, e.g., P.S. Farmer, Drug Design, Ariens, E.J., ed., Vol. 10, pp 119- 143 (Academic Press, New York, 1980); U.S. Patent No. 5,331,573; U.S. Patent No. 5,500,807; C. Verlinde, Structure. 2:577-587 (1994); and I. D. Kuntz, Science. 257:1078-1082 (1992). The model building techniques and computer evaluation systems described herein are not a limitation on the present invention. Thus, using these computer evaluation systems, a large number of compounds may be quickly and easily examined and expensive and lengthy biochemical testing avoided. Moreover, the need for actual synthesis of many compounds is effectively eliminated.
Once identified by the modelling techniques, the protease inhibitor may be tested for bioactivity using standard techniques. For example, structure of the invention may be used in binding assays using conventional formats to screen inhibitors. Suitable assays for use herein include, but are not limited to, the enzyme- linked immunosorbent assay (ELISA), or a fluoresence quench assay. See, for example, the cathepsin K activity assay of Example 2 below. Other assay formats may be used; these assay formats are not a limitation on the present invention.
In another aspect, the protease structure of the invention permit the design and identification of synthetic compounds and/or other molecules which have a shape complimentary to the conformation of the protease active site of the invention. Using known computer systems, the coordinates of the protease structure of the invention may be provided in machine readable form, the test compounds designed and/or screened and their conformations superimposed on the structure of the protease of the invention. Subsequently, suitable candidates identified as above may be screened for the desired protease inhibitory bioactivity, stability, and the like.
Once identified and screened for biological activity, these inhibitors may be used therapeutically or prophylactically to block cathepsin K activity.
The following examples illustrate various aspects of this invention. These examples do not limit the scope of this invention which is defined by the appended claims.
EXAMPLE 1 : Analysis of the Structure of procathepsin K
A. Expression, Purification and Crystallization
Plasmid constructions. Procathepsin K (procat K) was expressed using the PET system (Novagen). In order to express procat K, an existing clone containing the full-length human preprocathepsinK cDNA was subject to PCR amplifcation using oligos designed to introduce a Met residue at amino acid 18 of the full-length preproenzyme. The resulting fragment was cleaved with Ncol and
BamHI and this fragment was inserted into the pETlόb vector, resulting in pETprocat K.
Expression of recombinant. human procathepsin K. Isolation and solubilization of inclusion bodies. Recombinant pET vectors were transformed into strain BL21DE3 (Novagen). Bacteria were grown at 37C to OD6QO=0.6-0.7 and induced with ImM IPTG for 2 hours. The cell pellet from 6 L of bacterial culture weighing 15 g was washed with 150 mL of 50 mM Tris/HCl, 5 mM EDTA, 150 mM NaCl, pH 8.0. After centrifugation at 13,000 x g for 15 rnins, the washed pellet was resuspended into 150 mL of the same buffer prepared at 4 C and lysed by passage twice through a cell disruptor (Avestin) at 10,000 psi. The lysate was centrifuged as above, the supernatant decanted and the pellet suspended in 150 mL 50 mM Tris/HCl, 6 M guanidine HC1, 10 mM DTT, 5 mM EDTA, 150 mM NaCl, pH 8.0. After stirring for 1 hr at room temperature, insoluble cellular debris was removed by centrifugation at 23,000 x g for 30 mins and the supernatant clarified by filtration
(0.45 um PVDF, Millipore). The resulting solution was assayed for protein concentration according to the method of Bradford using the BioRad protein asay and bovine serum albumin obtained from Pierce as standard.
Refolding and Purification. 645 mg of procathepsin K was refolded by quick dilution of the denatured protein solution into 10 L of stirring, N2 (g) spraged 50 mM Tris/HCl, 5 mM EDTA, 10 mM reduced and 1 mM oxidized glutathione, 0.7 M L-arginine pH 8.0 prepared at room temperature. The solution was stirred overnight at 4° C then concentrated using tangential flow followed by a stirred cell (Amicon), both fitted with 10K membranes. The concentrate was clarified by centrifugation and filtration then dialyzed against 25 mM Na2PO4, 1.0
M NaCl, pH 7.0 (buffer A). The 44 mL dialysate containing 343 mg of protein was applied at a LFR= 23 cm/hr in two runs to a 2.6 x 90 cm column of Superdex 75
(Pharmacia) pre-equilibrated in buffer A. Fractions containing procathepsin K were identified and pooled based upon purity as observed on 15% acylamide SDS-PAGE gels prepared according to the method of Laemmli (Laemmli U.K., (1970) Nature
227, 690-685). Rainbow high molecular weight markers (Amersham) were used as standards. Gels were visualized with 0.25% Brilliant Blue R stain (Sigma). A portion of the 259 mg of protein recovered from the size exclusion columns was concentrated to 20 mg/mL using a centricon-10 then diluted 1:1 with 25 mM Na2PO4 pH 7.0 to yield a 10 mg/mL solution in 25 mM , 0.5 M NaCl, pH 7.0.
Crystals of procathepsin K grew to a size of approximately 0.3 mm^ in ten days at 4° C. The concentration of procathepsin K used in the crystallization was lOmg/ml. The hanging drops vapour diffusion method was used with a solution of 8-10% PEG6000, 0.1M Na citrate at pH 5.0. X-ray diffraction data were measured from a single crystal using a Siemens two-dimensional position-sensitive detector. Crystals of the complex are orthorhombic, space group C222ι , with cell constants of a=56.81 A, b=156.81 A, and c=96.10 A. The crystals contain one molecule in the asymmetric unit and contain 60% solvent.
B. Model Building and Refinement
The structure of procathepsin K was solved by molecular replacement using the program XPLOR (Brunger, A. T., Kuriyan, J. and Karplus, M. (1987) Science 235, 458-460). The coordinates for 217 residues from the structure of human cathespin K (Zhao, B. et al. (1977) Nature Struct. Biol. 4, 109-111) were used as a starting model. The cross rotation function was calculated using data from 10-3.5 A with a radius of integration of 23 A in a PI cell for the search model with dimensions of a=88.0 A, b=74.0 A, c=68.0 A. The search was carried out with pseudo-orthogonal euler angles in Patterson space from θl 0-180°,
Θ2 0-90, and Θ3 0-720°. The highest peak was 15.7 σ. The translation search was computed using data from 10 -3.5 A and gave a peak at 26 σ. The resulting model yielded a R-factor of 0.43. The R-factor for the model dropped to 0.40 after rigid body refinement from 10-3.5 A and to 0.31 after 200 cycles of positional refinement. The electron density maps were improved using solvent flattening and by histogram matching as implemented in the module DM of the CCP4 suite of programs (Collaborative Computational Project, Number 4 (1994), "The CCP4 Suite: Programs for Protein Crystallography", Acta Cryst. D50, 760-763). The modified electron density map was to build the model of used procathepsin K. Conventional positional refinement was used to refine the structure during model building. The structure was refined using X-plor. The final model of procathepsin K consists of residues 5 to 214. The simulated annealing refinement was finally carried out to a final R of 0.204. The backbone conformation of all residues is within the Ramachandran plot limits.
EXAMPLE 2: Assays
Determination of cathepsin K proteolytic catalytic activity
All assays for cathepsin K were carried out with human recombinant enzyme. Standard assay conditions for the determination of kinetic constants used a fluorogenic peptide substrate, typically Cbz-Phe-Arg-AMC, and were determined in 100 mM Na acetate at pH 5.5 containing 20 mM cysteine and 5 mM EDTA. Stock substrate solutions were prepared at concentrations of 10 or 20 mM in DMSO with 20 uM final substrate concentration in the assays. All assays contained 10% DMSO. Independent experiments found that this level of DMSO had no effect on enzyme activity or kinetic constants. All assays were conducted at ambient temperature. Product fluorescence (excitation at 360 nM; emission at 460 nM) was monitored with a Perceptive Biosystems Cytofluor II fluorescent plate reader. Product progress curves were generated over 20 to 30 minutes following formation of AMC product.
Inhibition studies
Potential inhibitors were evaluated using the progress curve method. Assays were carried out in the presence of variable concentrations of test compound. Reactions were initiated by addition of enzyme to buffered solutions of inhibitor and substrate. Data analysis was conducted according to one of two procedures depending on the appearance of the progress curves in the presence of inhibitors. For those compounds whose progress curves were linear, apparent inhibition constants (Ki>app) were calculated according to equation 1 (Brandt et al, Biochemistry, 1989, 28, 140):
v = VmA / [Ka(l + I/Kt> app) +A]
(1) where v is the velocity of the reaction with maximal velocity Vm , A is the concentration of substrate with Michaelis constant of Ka, and I is the concentration of inhibitor.
For those compounds whose progress curves showed downward curvature characteristic of time-dependent inhibition, the data from individual sets was analyzed to give k0bs according to equation 2:
[AMC] = vSs t + (vo - vss) [1 - exp (-kobst)J ' 'kσbs (2)
where [AMC] is the concentration of product formed over time t, vo is the initial reaction velocity and vss is the final steady state rate. Values for k0bs were then analyzed as a linear function of inhibitor concentration to generate an apparent second order rate constant (kDbs / inhibitor concentration or k0bs / [I]) describing the time-dependent inhibition. A complete discussion of this kinetic treatment has been fully described (Morrison et ah, Adv. Enzymol. Relat. Areas Mol. Biol., 1988, 61, 201).
This assay measures the affinity of inhibitors to cathepsin K. One skilled in the art would consider any compound exhibiting a Kj value of less than 50 micromolar to be a potential lead compound for further research. Human Osteoclast Resorption Assay
Aliquots of osteoclastoma-derived cell suspensions were removed from liquid nitrogen storage, warmed rapidly at 37°C and washed xl in RPMI-1640 medium by centrifugation (1000 rpm, 5 min at 4°C). The medium was aspirated and replaced with murine anti-HLA-DR antibody, diluted 1:3 in RPMI-1640 medium, and incubated for 30 min on ice The cell suspension was mixed frequently.
The cells were washed x2 with cold RPMI-1640 by centrifugation (1000 rpm, 5 min at 4°C) and then transferred to a sterile 15 mL centrifuge tube. The number of mononuclear cells were enumerated in an improved Neubauer counting chamber.
Sufficient magnetic beads (5 / mononuclear cell), coated with goat anti-mouse IgG, were removed from their stock bottle and placed into 5 mL of fresh medium (this washes away the toxic azide preservative). The medium was removed by immobilizing the beads on a magnet and is replaced with fresh medium. The beads were mixed with the cells and the suspension was incubated for 30 min on ice. The suspension was mixed frequently. The bead-coated cells were immobilized on a magnet and the remaining cells (osteoclast-rich fraction) were decanted into a sterile 50 mL centrifuge tube. Fresh medium was added to the bead- coated cells to dislodge any trapped osteoclasts. This wash process was repeated lO. The bead-coated cells were discarded.
The osteoclasts were enumerated in a counting chamber, using a large-bore disposable plastic Pasteur pipette to charge the chamber with the sample. The cells were pelleted by centrifugation and the density of osteoclasts adjusted to 1.5xlθ4/mL in EMEM medium, supplemented with 10% fetal calf serum and 1.7g/liter of sodium bicarbonate. 3 mL aliquots of the cell suspension ( per treatment) were decanted into 15 mL centrifuge tubes. These cells were pelleted by centrifugation. To each tube 3 mL of the appropriate treatment was added (diluted to 50 uM in the EMEM medium). Also included were appropriate vehicle controls, a positive control (87MEM1 diluted to 100 ug/mL) and an isotype control (IgG2a diluted to 100 ug/mL). The tubes were incubate at 37°C for 30 min.
0.5 mL aliquots of the cells were seeded onto sterile dentine slices in a 48- well plate and incubated at 37°C for 2 h. Each treatment was screened in quadruplicate. The slices were washed in six changes of warm PBS (10 mL / well in a 6-well plate) and then placed into fresh treatment or control and incubated at 37°C for 48 h. The slices were then washed in phosphate buffered saline and fixed in 2% glutaraldehyde (in 0.2M sodium cacodylate) for 5 min., following which they were washed in water and incubated in buffer for 5 min at 37°C. The slices were then washed in cold water and incubated in cold acetate buffer / fast red garnet for 5 min at 4°C. Excess buffer was aspirated, and the slices were air dried following a wash in water.
The TRAP positive osteoclasts were enumerated by bright-field microscopy and were then removed from the surface of the dentine by sonication. Pit volumes were determined using the Nikon/Lasertec ILM21W confocal microscope.
EXAMPLE 3: Method of Detecting Inhibitors
The three dimensional atomic structure can be readily used as a template for selecting potent inhibitors. Various computer programs and databases are available for the purpose. A good inhibitor should at least have excellent steric and electrostatic complementarity to the target, a fair amount of hydrophobic surface buried and sufficent conformational rigidity to minimize entropy loss upon binding. The approach usually comprises several steps: 1) Define a region to target, the active site cavity of procathepsin K can be selected, but any place that is essential to the protease activity could become a potential target. Since the crystal structure has been determined, the spatial and chemical properties of the target region is known.
2) Docking a small molecule onto the target. Many methods can be used to archive this. Computer databases of three-dimensional structures are available for screening millions of small molecular compounds. A negative image of these compounds can be calculated and used to match the shape of the target cavity. The profiles of hydrogen bond donor-acceptor and lipophilic points of these compounds can also be used to complement those of the target. Anyone skilled in the art would be able to indentify many small molecules or fragments as hits. 3) Linking and extending recognzition fragments. Using the hits identified by above procedure, one can incoφorate different functional groups or small molecules into a single, larger molecule. The resulting molecule is likely to be more potent and have higher specificity. It is also possible to try to improve the "seed" inhibitor by adding more atoms or fragments that will interact with the target protein. The originally defined target region can be readily expanded to allow further necessary extension.
A limited numer of promising compounds can be selected through the process. They can then be synthesized and assayed for their inhibitory properties. The success rate can sometimes be as high as 20%, and it may still be higher with the rapid progresses in computing methods.
AbbrevaiationsE-64. [ 1 -[N-[(L-3-tr n_-carboxyoxirane-2carbonyl)-
L-leucyl]amino]-4-guanidinobutane]
CBZ, benzyloxycarbonyl AMC, aminomethylcoumarin
MPD, 2 methyl-2,4-pentanediol
PIPES, piperazone-N,N-bis(2-ethanesulfonic acid)
MES, 2-(N-morpholino)-ethanesulfonic acid tris, tris(hydroxymethyl)-aminomethane PEG, polyethyleneglycol
M, Molar
RC = ΣI(F0 - FC)I / F0
F0 = observed structure amplitude
Fc = calculated structure amplitude EDTA, ethylenediaminetetraacetic acid DTT, 1,4-dithiothreitol SDS-PAGE, sodium dodecylsulfate polyacrylamide gel electrophoresis This invention is not to be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.
The disclosures of the patents, patent applications and publications cited herein are incorporated by reference in their entireties.
Table I Table of the orthogonal three dimensional coordinates in Angstroms and B factors (A2) for procathepsin K.
Residue Atom X Y Z B
5 GLU CB 10, .66 -1, .63 54 .79 15, .00
5 GLU CG 10. .16 -2, .56 53 .66 15, .00
5 GLU CD 9, .13 -3. .60 54 .12 15, .00
5 GLU OE1 8, .82 -3. .66 55, .34 15, .00
5 GLU OE2 8. .63 -4. .36 53, .25 15. .00
5 GLU C 11. .61 -1. .76 .57, .08 15. .00
5 GLU 0 11, .20 -2. .60 57, .88 15, .00
5 GLU N 11 .93 -3. .64 55 .60 15, .00
5 GLU CA 11 .84 -2, .15 55 .63 15, ,00
6 ILE N 11, .90 -0, .50 57, .39 15. .00
6 ILE CA 11, .75 0, .09 58, .73 15. .00
6 ILE CB 12, .62 -0. .64 59, .80 15, .00
6 ILE CG2 11, .83 -1. .76 60, .47 15. ,00
6 ILE CGI 13, .96 -1. .08 59, .19 15. ,00
6 ILE CD1 14, .78 -2, .00 60, .08 15. .00
6 ILE C 12, .20 1, .55 58, .67 15. ,00
6 ILE 0 12, .53 2. .17 59, .70 15. .00
7 LEU N 12, .15 2. .12 57, .47 15. ,00
7 LEU CA 12. .57 3. .50 57. .23 15. ,00
7 LEU CB 12. .18 3. .97 55. .84 15. ,00
7 LEU CG 12. .66 3, .11 .54, .68 15. .00
7 LEU CD1 12, .56 3, .91 53. .40 15. ,00
7 LEU CD2 14, .09 2. .67 54. .90 15. ,00
7 LEU C 12, .09 4. .50 58, .26 15. ,00
7 LEU 0 12. .76 5. .49 58. .49 15. ,00
8 ASP N 10. .95 4. .24 58. .89 15. .00
8 ASP CA 10. .46 5. .19 59. .88 15. ,00
8 ASP CB 9, .09 4. .80 60, .45 15. ,00
8 ASP CG 8. .27 6. .03 60, .89 15. .00
8 ASP OD1 7. .82 6. .81 60. .01 15. ,00
8 ASP OD2 8. .09 6. .23 62. .12 15. ,00 ASP C 11.48 5.28 61.00 15.00 ASP 0 11.95 6.38 61.35 15.00 THR N 11.91 4.13 61.50 15.00 THR CA 12.88 4.16 62.57 15.00 THR CB 13.03 2.79 63.24 15.00 THR OG1 13.87 2.91 64.39 15.00 THR CG2 13.60 1.76 62.26 15.00 THR C 14.24 4.69 62.10 15.00 THR 0 14.83 5.57 62.74 15.00 HIS N 14.69 4.22 60.94 15.00 HIS CA 15.97 4.65 60.41 15.00 HIS CB 16.36 3.81 59.21 15.00 HIS CG 16.86 2.47 59.60 15.00 HIS CD2 17.15 1.96 60.82 15.00 HIS NDl 17.15 1.48 58.69 15.00 HIS CEl 17.60 0.41 59.34 15.00 HIS NE2 17.61 0.68 60.63 15.00 HIS C 16.15 6.12 60.11 15.00 HIS 0 17.18 6.68 60.45 15.00 TRP N 15.14 6.74 59.50 15.00 TRP CA 15.23 8.16 59.19 15.00 TRP CB 13.96 8.67 58.51 15.00 TRP CG 14.17 9.99 57.80 15.00 TRP CD2 13.47 11.22 58.04 15.00 TRP CE2 14.00 12.18 57.14 15.00 TRP CE3 12.46 11.60 58.93 15.00 TRP CD1 15.06 10.25 56.79 15.00 TRP NE1 14.97 11.56 56.39 15.00 TRP CZ2 13.54 13.50 57.10 15.00 TRP CZ3 12.00 12.91 58.89 15.00 TRP CH2 12.54 13.84 57.98 15.00 TRP C 15.47 8.94 60.48 15.00 TRP 0 16.18 9.94 60.48 15.00 GLU N 14.93 8.43 61.58 15.00 GLU CA 15.11 9.06 62.88 15.00 GLU CB 14.29 8.34 63.95 15.00 GLU CG 14.34 9.01 65.32 15.00 GLU CD 13.63 10.36 65.38 15.00 GLU OEl 13.03 10.80 64.36 15.00 GLU OE2 13.66 10.98 66.47 15.00 GLU C 16.58 9.07 63.27 15.00 GLU 0 17.03 9.94 64.01 15.00 LEU N 17.32 8.07 '62.81 15.00 LEU CA 18.75 8.02 63.10 15.00 LEU CB 19.33 6.62 62.88 15.00 LEU CG 20.35 6.13 63.92 15.00 LEU CDl 21.11 4.94 63.36 15.00 LEU CD2 21.33 7.24 64.32 15.00 LEU C 19.39 9.03 62.16 15.00 LEU 0 19.84 10.08 62.59 15.00 TRP N 19.34 8.75 60.85 15.00 TRP CA 19.91 9.62 59.81 15.00 TRP CB 19.27 9.31 58.46 15.00 TRP CG 19.90 10.01 57.31 15.00 TRP CD2 19.67 11.36 56.89 15.00 TRP CE2 20.40 11.55 55.71 15.00 TRP CE3 18.90 12.42 57.39 15.00 TRP CDl 20.75 9.47 '56.41 15.00 TRP NE1 21.06 10.38 55.43 15.00 TRP CZ2 20.40 12.77 55.02 15.00 TRP CZ3 18.90 13.63 56.70 15.00 TRP CH2 19.64 13.79 55.53 15.00 TRP C 19.72 11.10 60.11 15.00 TRP 0 20.60 11.91 59.82 15.00 LYS N 18.55 11.46 60.63 15.00 LYS CA 18.27 12.83 60.97 15.00 LYS CB 16.86 12.96 61.54 15.00 LYS CG 15.81 13.43 60.53 15.00 LYS CD 14.44 13.50 61.18 15.00 LYS CE 14.51 14.18 62.55 15.00 LYS NZ 15.05 15.57 62.48 15.00 LYS C 19.27 13.25 62.04 15.00 LYS 0 20.15 14.10 '61.80 15.00 LYS N 19.19 12.58 63.18 15.00 LYS CA 20.05 12.86 64.31 15.00 LYS CB 19.56 12.12 65.55 15.00 LYS CG 19.67 10.62 65.47 15.00 LYS CD 18.71 9.97 66.46 15.00 LYS CE 18.98 8.49 66.61 15.00 LYS NZ 20.38 8.24 67.09 15.00 LYS C 21.52 12.55 64.09 15.00 LYS 0 22.37 13.10 64.78 15.00 THR N 21.82 11.67 63.14 15.00 THR CA 23.20 11.31 .62.85 15.00 THR CB 23.36 10.18 61.82 15.00 THR OGl 22.32 9.22 61.98 15.00 THR CG2 24.71 9.49 62.02 15.00 THR C 23.85 12.54 62.25 15.00 THR 0 25.07 12.65 62.18 15.00 HIS N 23.03 13.44 61.73 15.00 HIS CA 23.55 14.65 61.16 15.00 HIS CB 23.13 14.78 59.70 15.00 HIS CG 23.71 13.71 58.82 15.00 HIS CD2 24.64 13.77 57.84 15.00 HIS NDl 23.32 12.40 58.90 15.00 HIS CEl 23.99 11.69 58.01 15.00 HIS NE2 24.80 12.50 57.35 15.00 HIS C 22.98 15.77 62.02 15.00 HIS 0 22.58 15.51 •63.16 15.00 ARG N 22.98 16.99 61.51 15.00 ARG CA 22.45 18.13 62.26 15.00 ARG CB 23.41 19.33 62.19 15.00 ARG CG 24.11 19.55 60.83 15.00 ARG CD 25.62 19.36 60.95 15.00 ARG NE 26.28 20.55 61.49 15.00 ARG CZ 27.60 20.71 61.62 15.00 ARG NHl 28.08 21.85 62.10 15.00 ARG NH2 28.44 19.71 61.32 15.00 ARG C 21.05 18.50 61.74 15.00 ARG 0 20.65 19.67 61.76 15.00 LYS N 20.32 17.49 61.27 15.00 LYS CA 18.99 17.69 60.74 15.00 LYS CB 18.53 16.48 59.93 15.00 LYS CG 19.50 16.05 58.86 15.00 LYS CD 19.70 17.13 57.84 15.00 LYS CE 20.67 16.65 56.78 15.00 LYS NZ 20.71 17.52 55.56 15.00 LYS C 18.02 17.91 61.89 15.00 LYS 0 17.88 17.06 62.78 15.00 GLN N 17.42 19.09 61.90 15.00 GLN CA 16.45 19.47 62.91 15.00 GLN CB 17.16 20.10 64.12 15.00 GLN CG 16.35 20.08 65.40 15.00 GLN CD 16.09 18.67 65.97 15.00 GLN OEl 15.74 18.53 67.14 15.00 GLN NE2 16.27 17.63 '65.14 15.00 GLN C 15.69 20.49 62.11 15.00 GLN 0 16.13 21.63 61.96 15.00 TYR N 14.62 20.02 61.47 15.00 TYR CA 13.80 20.86 60.60 15.00 TYR CB 12.98 20.00 59.65 15.00 TYR CG 13.82 18.98 58.92 15.00 TYR CDl 14.37 17.89 59.59 15.00 TYR CEl 15.08 16.91 58.91 15.00 TYR CD2 14.02 19.07 57.54 15.00 TYR CE2 14.72 18.10 56.86 15.00 TYR CZ 15.25 17.02 57.55 15.00 TYR OH 15.92 16.05 56.86 15.00 TYR C 12.88 21.73 61.43 15.00 TYR 0 12.54 21.37 62.56 15.00 ASN N 12.48 22.87 '60.88 15.00 ASN CA 11.62 23.77 61.63 15.00 ASN CB 11.83 25.24 61.22 15.00 ASN CG 11.47 25.52 59.76 15.00 ASN OD1 10.44 25.07 59.26 15.00 ASN ND2 12.34 26.26 59.07 15.00 ASN C 10.13 23.43 61.61 15.00 ASN 0 9.34 24.13 62.25 15.00 ASN N 9.76 22.35 60.91 15.00 ASN CA 8.35 21.95 60.81 15.00 ASN CB 7.55 23.04 60.11 15.00 ASN CG 6.49 22.48 .59.19 15.00 ASN ODl 5.48 21.92 59.65 15.00 ASN ND2 6.73 22.57 57.89 15.00 ASN C 8.11 20.62 60.07 15.00 ASN 0 8.84 20.29 59.13 15.00 LYS N 7.00 19.96 60.40 15.00 LYS CA 6.62 18.68 59.79 15.00 LYS CB 5.46 18.01 60.53 15.00 LYS CG 5.87 17.19 61.76 15.00 LYS CD 5.71 17.97 63.06 15.00 LYS CE 6.32 17.20 64.25 15.00 LYS NZ 7.84 17.31 64.32 15.00 LYS C 6.29 18.75 58.31 15.00 LYS 0 6.55 17.81 57.58 15.00 VAL N 5.67 19.84 57.86 15.00 VAL CA 5.34 19.92 .56.44 15.00 VAL CB 4.57 21.24 56.06 15.00 VAL CGI 4.44 21.39 54.55 15.00 VAL CG2 3.17 21.22 56.68 15.00 VAL C 6.66 19.78 55.68 15.00 VAL 0 6.74 19.01 54.72 15.00 ASP N 7.71 20.44 56.17 15.00 ASP CA 9.02 20.33 55.53 15.00 ASP CB 10.04 21.31 56.10 15.00 ASP CG 9.46 22.65 56.38 15.00 ASP ODl 9.75 23.18 57.48 15.00 ASP OD2 8.71 23.18 55.52 15.00 ASP C 9.50 18.93 55.84 15.00 ASP 0 9.69 18.12 54.93 15.00 GLU N 9.65 18.64 57.13 15.00 GLU CA 10.12 17.34 .57.58 15.00 GLU CB 9.66 17.07 59.01 15.00 GLU CG 10.34 15.87 59.63 15.00 GLU CD 10.44 15.93 61.15 15.00 GLU OEl 10.03 16.95 61.74 15.00 GLU OE2 10.94 14.95 61.74 15.00 GLU C 9.64 16.24 56.65 15.00 GLU 0 10.43 15.68 55.89 15.00 ILE N 8.33 16.04 56.59 15.00 ILE CA 7.73 15.02 55.74 15.00 ILE CB 6.16 15.07 55.73 15.00 ILE CG2 5.60 13.77 55.16 15.00 ILE CGI 5.56 15.32 •57.13 15.00 ILE CDl 6.01 14.36 58.22 15.00 ILE C 8.20 15.19 54.29 15.00 ILE 0 8.50 14.21 53.62 15.00 SER N 8.29 16.44 53.83 15.00 SER CA 8.69 16.70 52.44 15.00 SER CB 8.47 18.17 52.07 15.00 SER OG 9.56 18.98 52.47 15.00 SER C 10.13 16.33 52.18 15.00 SER 0 10.44 15.77 51.12 15.00 ARG N 11.02 16.66 53.12 15.00 ARG CA 12.45 16.36 52.99 15.00 ARG CB 13.21 16.98 54.15 15.00 ARG CG 12.99 18.47 54.28 15.00 ARG CD 14.11 19.29 53.67 15.00 ARG NE 14.07 19.35 '52.21 15.00 ARG CZ 14.49 20.39 51.51 15.00 ARG NHl 14.98 21.46 52.13 15.00 ARG NH2 14.44 20.38 50.18 15.00 ARG C 12.57 14.85 53.05 15.00 ARG 0 13.27 14.24 52.25 15.00 ARG N 11.81 14.27 53.97 15.00 ARG CA 11.75 12.83 54.16 15.00 ARG CB 10.79 12.54 55.31 15.00 ARG CG 10.21 11.16 55.32 15.00 ARG CD 11.27 10.14 55.59 15.00 ARG NE 10.70 8.80 55.66 15.00 ARG CZ 10.00 8.36 56.70 15.00 ARG NHl 9.79 9.14 57.76 15.00 ARG NH2 9.48 7.13 56.66 15.00 ARG C 11.29 12.12 52.88 15.00 ARG 0 11.68 10.98 52.63 15.00 LEU N 10.47 12.78 52.08 15.00 LEU CA 10.01 12.18 50.83 15.00 LEU CB 8.74 12.85 50.31 15.00 LEU CG 7.83 11.79 49.70 15.00 LEU CDl 6.64 11.61 50.61 15.00 LEU CD2 7.42 12.15 48.27 15.00 LEU C 11.10 12.25 49.79 15.00 LEU 0 11.16 11.41 48.90 15.00 ILE N 11.93 13.29 49.88 15.00 ILE CA 13.05 13.43 48.96 15.00 ILE CB 13.71 14.81 49.07 15.00 ILE CG2 14.67 14.99 47.92 15.00 ILE CGI 12.67 15.92 48.96 15.00 ILE CDl 13.24 17.31 49.09 15.00 ILE C 14.07 12.36 49.36 15.00 ILE 0 14.56 11.59 48.52 15.00 TRP N 14.32 12.27 50.66 15.00 TRP CA 15.25 11.29 51.22 15.00 TRP CB 15.08 11.24 52.73 15.00 TRP CG 15.93 10.24 53.44 15.00 TRP CD2 15.53 8.95 53.90 15.00 TRP CE2 16.58 8.45 54.70 15.00 TRP CE3 14.38 8.17 53.73 15.00 TRP CDl 17.18 10.45 53.94 15.00 TRP NEl 17.57 9.39 54.70 15.00 TRP CZ2 16.51 7.21 55.33 15.00 TRP CZ3 14.32 6.93 54.37 15.00 TRP CH2 15.37 6.47 55.16 15.00 TRP C 14.91 9.94 50.64 15.00 TRP 0 15.78 9.22 50.15 15.00 GLU N 13.62 9.62 50.66 15.00 GLU CA 13.12 8.37 50.15 15.00 GLU CB 11.65 8.21 50.51 15.00 GLU CG 11.38 8.22 52.01 15.00 GLU CD 10,.29 7..27 52.42 15.00
GLU OEl 9, .95 7. .27 53 .62 15, .00
GLU OE2 9, .79 6, .50 •51, .56 15, .00
GLU C 13, .33 8. .22 48 .65 15, .00
GLU 0 13, .63 7 .13 48 .17 15, .00
LYS N 13, .17 9, .30 47, .90 15. .00
LYS CA 13. .37 9 .25 46 .45 15, .00
LYS CB 13, .06 10, .59 45, .79 15, .00
LYS CG 11, .56 10 .88 45 .66 15, .00
LYS CD 11, .27 12 .10 44 .78 15, .00
LYS CE 11, .74 13, .38 45, .46 15. .00
LYS NZ 11, .14 13 .54 46 .83 15. .00
LYS C 14, .80 8. .83 46, .13 15. .00
LYS 0 15. .03 7, .80 45, .48 15, .00
ASN N 15, .76 9 .61 46 .66 15. .00
ASN CA 17. .19 9, .38 46, .48 15. .00
ASN CB 18, .00 10 .45 47, .19 15. .00
ASN CG 17, .81 11, .81 •46, .58 15, .00
ASN ODl 17. .65 11, .94 45, .36 15. ,00
ASN ND2 17, .84 12 .84 47, .42 15, ,00
ASN C 17. .63 8, .02 46, .98 15. .00
ASN 0 18. .41 7, .34 46, .31 15. ,00
LEU N 17, .18 7, .64 48, .18 15, ,00
LEU CA 17. .53 6, .34 48, .76 15. .00
LEU CB 16, .73 6, .07 50, .05 15, ,00
LEU CG 17. .36 5, .16 51, .11 15. .00
LEU CDl 16. .40 4, ,97 52. .27 15. .00
LEU CD2 17. .71 3, .80 50. .53 15. .00
LEU C 17, .15 5, .30 47. .71 15. ,00
LEU 0 17. .96 4, ,44 47. ,33 15. ,00
LYS N 15, ,95 5, .46 47. ,18 15. ,00
LYS CA 15, ,43 4. .57 46. ,16 15. ,00
LYS CB 13. .99 4, .99 '45. ,83 15. ,00
LYS CG 13, .13 3. .90 45, .22 15. ,00
LYS CD 11. ,66 4, .08 45. .59 15. 00
LYS CE 11. ,12 5, .44 45. ,17 15. ,00
LYS NZ 11. ,27 5, .65 43. ,70 15. 00 LYS C 16.34 4.65 44.94 15.00 LYS 0 16.71 3.63 44.36 15.00 TYR N 16.75 5.87 44.60 15.00 TYR CA 17.60 6.11 43.44 15.00 TYR CB 17.90 7.61 43.27 15.00 TYR CG 18.84 7.87 42.12 15.00 TYR CDl 20.14 8.32 42.34 15.00 TYR CEl 21.06 8.39 .41.28 15.00 TYR CD2 18.46 7.53 40.82 15.00 TYR CE2 19.36 7.60 39.76 15.00 TYR CZ 20.66 8.02 39.99 15.00 TYR OH 21.56 7.98 38.95 15.00 TYR C 18.92 5.36 43.57 15.00 TYR 0 19.40 4.76 42.61 15.00 ILE N 19.49 5.42 44.76 15.00 ILE CA 20.76 4.77 45.03 15.00 ILE CB 21.22 5.07 46.46 15.00 ILE CG2 22.50 4.32 46.76 15.00 ILE CGI 21.45 6.57 46.63 15.00 ILE CDl 21.94 6.97 47.99 15.00 ILE C 20.63 3.28 44.85 15.00 ILE 0 21.25 2.71 43.97 15.00 SER N 19.75 2.68 .45.64 15.00 SER CA 19.48 1.25 45.64 15.00 SER CB 18.16 0.97 46.38 15.00 SER OG 17.89 -0.41 46.51 15.00 SER C 19.46 0.60 44.26 15.00 SER 0 19.91 -0.54 44.11 15.00 ILE N 18.98 1.31 43.26 15.00 ILE CA 18.91 0.79 41.90 15.00 ILE CB 17.78 1.47 41.08 15.00 ILE CG2 17.85 1.04 39.63 15.00 ILE CGI 16.41 1.14 41.69 15.00 ILE CDl 15.33 2.08 41.25 15.00 ILE C 20.22 0.99 41.17 15.00 ILE 0 20.65 0.13 40.40 15.00 HIS N 20.82 2.16 41.37 15.00 HIS CA 22.09 2.49 40.73 15.00 HIS CB 22.54 3.90 41.14 15.00 HIS CG 23.66 4.46 40.30 15.00 HIS CD2 23.65 5.22 39.19 15.00 HIS NDl 25.00 4.24 40.60 15.00 HIS CEl 25.75 4.86 39.70 15.00 HIS NE2 24.96 5.46 38.84 15.00 HIS C 23.11 1.43 41.16 15.00 HIS 0 23.80 0.85 40.32 15.00 ASN N 23.18 1.15 42.45 15.00 ASN CA 24.11 0.14 42.93 15.00 ASN CB 24.06 0.01 44.45 15.00 ASN CG 24.80 1.13 45.15 15.00 ASN ODl 25.23 2.09 44.52 15.00 ASN ND2 24.95 1.01 46.44 15.00 ASN C 23.77 -1.19 42.26 15.00 ASN 0 24.65 -1.87 41.75 15.00 LEU N 22.48 -1.55 42.22 15.00 LEU CA 22.07 -2.79 41.58 15.00 LEU CB 20.53 -2.86 41.50 15.00 LEU CG 19.85 -4.07 40.82 15.00 LEU CDl 20.08 -5.34 41.62 15.00 LEU CD2 18.36 -3.80 40.71 15.00 LEU C 22.66 -2.80 40.17 15.00 LEU 0 23.32 -3.76 39.79 15.00 GLU N 22.48 -1.71 39.43 15.00 GLU CA 23.01 -1.62 38.07 15.00 GLU CB 22.68 -0.26 37.42 15.00 GLU CG 21.19 0.09 37.26 15.00 GLU CD 20.95 1.39 36.46 15.00 GLU OEl 20.57 2.43 37.06 15.00 GLU OE2 21.13 1.37 35.22 15.00 GLU C 24.52 -1.84 38.09 15.00 GLU 0 25.08 -2.40 37.14 15.00 ALA N 25.18 -1.41 39.16 15.00 ALA CA 26.64 -1.57 39.31 15.00 ALA CB 27.18 -0.75 40.48 15.00 ALA C 27.03 -3.04 39.48 15.00 ALA 0 28.07 -3.45 .38.98 15.00 SER N 26.22 -3.81 40.19 15.00 SER CA 26.48 -5.23 40.38 15.00 SER CB 25.41 -5.86 41.28 15.00 SER OG 24.70 -4.87 42.02 15.00 SER C 26.43 -5.91 39.02 15.00 SER 0 26.98 -6.99 38.81 15.00 LEU N 25.69 -5.29 38.10 15.00 LEU CA 25.58 -5.82 36.76 15.00 LEU CB 24.17 -5.61 36.20 15.00 LEU CG 23.04 -6.48 36.74 15.00 LEU CDl 23.53 -7.90 36.84 15.00 LEU CD2 22.54 -5.99 38.09 15.00 LEU C 26.63 -5.11 35.91 15.00 LEU 0 26.61 -5.15 34.67 15.00 GLY N 27.51 -4.39 .36.60 15.00 GLY CA 28.60 -3.68 35.96 15.00 GLY C 28.37 -2.64 34.88 15.00 GLY 0 29.30 -2.32 34.15 15.00 VAL N 27.16 -2.11 34.77 15.00 VAL CA 26.91 -1.09 33.74 15.00 VAL CB 25.43 -1.05 33.33 15.00 VAL CGI 25.06 -2.34 32.62 15.00 VAL CG2 24.54 -0.83 34.53 15.00 VAL C 27.42 0.30 34.15 15.00 VAL 0 27.41 1.24 33.35 15.00 HIS N 27.88 0.41 35.40 15.00 HIS CA 28.43 1.66 35.94 15.00 HIS CB 27.55 2.19 37.06 15.00 HIS CG 26.18 2.62 36.61 15.00 HIS CD2 25.05 2.86 .37.30 15.00 HIS NDl 25.87 2.83 35.29 15.00 HIS CEl 24.60 3.18 35.18 15.00 HIS NE2 24.08 3.21 36.40 15.00 HIS C 29.79 1.27 36.51 15.00 HIS 0 30.05 0.07 36.66 15.00 THR N 30.69 2.23 36.76 15.00 THR CA 31.98 1.87 37.34 15.00 THR CB 33.17 2.13 36.41 15.00 THR OGl 33.23 3.51 36.08 15.00 THR CG2 33.04 1.30 35.15 15.00 THR C 32.23 2.55 38.68 15.00 THR 0 33.37 2.63 '39.15 15.00 TYR N 31.14 3.00 39.29 15.00 TYR CA 31.17 3.67 40.59 15.00 TYR CB 31.14 5.18 40.44 15.00 TYR CG 30.10 5.69 39.48 15.00 TYR CDl 29.01 6.42 39.93 15.00 TYR CEl 28.06 6.87 39.05 15.00 TYR CD2 30.21 5.42 38.12 15.00 TYR CE2 29.26 5.86 37.23 15.00 TYR CZ 28.18 6.58 37.70 15.00 TYR OH 27.23 6.98 36.80 15.00 TYR C 29.95 3.20 41.36 15.00 TYR 0 29.05 2.58 40.80 15.00 GLU N 29.93 3.51 42.65 15.00 GLU CA 28.84 3.12 43.53 15.00 GLU CB 29.25 1.87 44.31 15.00 GLU CG 29.73 0.76 43.38 15.00 GLU CD 30.28 -0.45 44.10 15.00 GLU OEl 30.16 -1.56 43.52 15.00 GLU OE2 30.84 -0.30 45.21 15.00 GLU C 28.51 4.29 44.46 15.00 GLU 0 29.41 5.06 44.85 15.00 LEU N 27.22 4.43 44.79 15.00 LEU CA 26.71 5.51 45.62 15.00 LEU CB 25.41 6.03 45.03 15.00 LEU CG 25.33 6.69 43.65 15.00 LEU CDl 26.48 6.32 42.76 15.00 LEU CD2 24.02 6.29 43.02 15.00 LEU C 26.42 5.08 47.04 15.00 LEU 0 26.37 3.88 47.34 15.00 ALA N 26.18 6.06 47.91 15.00 ALA CA 25.85 5.82 49.31 15.00 ALA CB 27.08 5.49 50.11 15.00 ALA C 25.16 7.07 49.86 15.00 ALA 0 25.24 8.13 49.25 15.00 MET N 24.48 6.95 50.98 15.00 MET CA 23.75 8.07 51.56 15.00 MET CB 22.55 7.54 52.34 15.00 MET CG 21.83 8.51 53.27 15.00 MET SD 20.90 9.80 52.48 15.00 MET CE 20.32 8.94 51.04 15.00 MET C 24.62 9.01 52.40 15.00 MET 0 25.05 8.69 53.50 15.00 ASN N 24.85 10.20 51.85 15.00 ASN CA 25.66 11.22 52.49 15.00 ASN CB 26.54 11.87 51.42 15.00 ASN CG 25.73 12.41 50.29 15.00 ASN ODl 24.80 13.17 50.50 15.00 ASN ND2 26.05 11.98 49.08 15.00 ASN C 24.78 12.29 53.16 15.00 ASN 0 23.56 12.26 53.02 15.00 HIS N 25.40 13.25 53.86 15.00 HIS CA 24.64 14.31 54.55 15.00 HIS CB 25.58 15.21 55.37 15.00 HIS CG 26.57 16.01 54.57 15.00 HIS CD2 27.33 17.08 54.91 15.00 HIS NDl 26.89 15.72 53.26 15.00 HIS CEl 27.81 16.57 52.83 15.00 HIS NE2 28.09 17.41 53.82 15.00 HIS C 23.78 15.19 53.63 15.00 HIS 0 23.40 16.30 54.01 15.00 LEU N 23.51 14.73 52.41 15.00 LEU CA 22.71 15.49 51.47 15.00 LEU CB 23.58 15.96 50.30 15.00 LEU CG 24.91 16.63 50.64 15.00 LEϋ CDl 25.52 17.18 49.38 15.00 LEU CD2 24.70 17.72 51.64 15.00 LEU C 21.56 14.61 50.98 15.00 LEU 0 21.18 14.65 •49.81 15.00 GLY N 21.02 13.79 51.87 15.00 GLY CA 19.91 12.93 51.52 15.00 GLY C 18.67 13.77 51.25 15.00 GLY 0 18.14 13.73 50.14 15.00 ASP N 18.27 14.57 52.23 15.00 ASP CA 17.08 15.43 52.13 15.00 ASP CB 16.64 15.92 53.51 15.00 ASP CG 17.70 16.74 54.24 15.00 ASP ODl 17.88 16.50 55.45 15.00 ASP OD2 18.34 17.66 53.66 15.00 ASP C 17.23 16.61 51.19 15.00 ASP 0 16.84 17.71 51.52 15.00 MET N 17.74 16.38 50.00 15.00 MET CA 17.93 17.45 49.05 15.00 MET CB 19.28 18.16 •49.28 15.00 MET CG 19.43 18.98 50.57 15.00 MET SD 20.95 20.02 50.58 15.00 MET CE 21.46 19.92 52.32 15.00 MET C 17.97 16.77 47.70 15.00 MET 0 17.69 15.58 47.60 15.00 THR N 18.35 17.51 46.67 15.00 THR CA 18.49 16.96 45.34 15.00 THR CB 17.13 16.84 44.60 15.00 THR OGl 17.35 16.31 43.28 15.00 THR CG2 16.43 18.20 44.52 15.00 THR C 19.45 17.82 44.52 15.00 THR 0 19.75 18.96 44.90 15.00 SER N 19.88 17.27 43.40 15.00 SER CA 20.80 17.91 42.47 15.00 SER CB 20.81 17.13 •41.17 15.00 SER OG 19.54 16.53 40.96 15.00 SER C 20.58 19.39 42.18 15.00 SER 0 21.54 20.14 42.07 15.00 GLU N 19.33 19.81 42.03 15.00 GLU CA 19.05 21.22 41.75 15.00 GLU CB 17.59 21.44 41.33 15.00 GLU CG 17.26 22.91 41.02 15.00 GLU CD 15.77 23.27 41.20 15.00 GLU OEl 15.47 24.38 41.69 15.00 GLU OE2 14.90 22.44 40.84 15.00 GLU C 19.36 22.06 42.98 15.00 GLU 0 19.78 23.21 42.87 15.00 GLU N 19.20 21.48 44.17 15.00 GLU CA 19.46 22.19 45.41 15.00 GLU CB 18.66 21.56 46.53 15.00 GLU CG 17.18 21.54 46.17 15.00 GLU CD 16.27 21.05 47.26 15.00 GLU OEl 16.05 21.80 48.24 15.00 GLU OE2 15.74 19.92 47.11 15.00 GLU C 20.94 22.31 45.73 15.00 GLU 0 21.44 23.40 46.03 15.00 VAL N 21.67 21.21 45.62 15.00 VAL CA 23.10 21.24 45.89 15.00 VAL CB 23.78 19.86 45.70 15.00 VAL CGI 23.19 18.83 46.65 15.00 VAL CG2 23.64 19.39 44.27 15.00 VAL C 23.70 22.24 44.93 15.00 VAL 0 24.53 23.06 45.31 15.00 VAL N 23.24 22.21 43.69 15.00 VAL CA 23.75 23.12 42.70 15.00 VAL CB 23.34 22.69 41.28 15.00 VAL CGI 23.70 23.75 40.28 15.00 VAL CG2 24.02 21.39 40.91 15.00 VAL C 23.27 24.53 42.99 15.00 VAL 0 23.97 25.48 42.68 15.00 GLN N 22.12 24.66 43.64 15.00 GLN CA 21.58 26.00 43.92 15.00 GLN CB 20.07 25.95 44.18 15.00 GLN CG 19.40 27.35 44.23 15.00 GLN CD 18.27 27.46 45.24 15.00 GLN OEl 17.55 26.49 45.51 15.00 GLN NE2 18.11 28.65 45.83 15.00 GLN C 22.24 26.70 45.09 15.00 73 GLN 0 22.43 27.90 45.05 15.00
74 LYS N 22.55 25.96 46.15 15.00
74 LYS CA 23.15 26.60 47.31 15.00
74 LYS CB 22.15 26.59 48.48 15.00
74 LYS CG 21.46 25.25 48.73 15.00
74 LYS CD 20.41 25.32 49.88 15.00
74 LYS CE 19.11 26.03 49.46 15.00
74 LYS NZ 18.34 25.38 48.33 15.00
74 LYS C 24.53 26.07 47.77 15.00
74 LYS 0 25.00 26.43 .48.86 15.00
75 MET N 25.19 25.27 46.93 15.00
75 MET CA 26.49 24.70 47.29 15.00
75 MET CB 26.30 23.34 47.96 15.00
75 MET CG 25.56 23.38 49.25 15.00
75 MET SD 25.38 21.74 49.87 15.00
75 MET CE 26.23 21.91 51.38 15.00
75 MET C 27.52 24.57 46.17 15.00
75 MET 0 28.31 23.63 46.18 15.00
76 THR N 27.50 25.45 45.19 15.00
76 THR CA 28.48 25.45 44.12 15.00
76 THR CB 27.97 24.75 42.84 15.00
76 THR OGl 26.90 25.50 42.28 15.00
76 THR CG2 27.50 23.34 43.14 15.00
76 THR C 28.74 26.93 43.86 15.00
76 THR 0 28.92 27.68 ■44.82 15.00
77 GLY N 28.75 27.36 42.60 15.00
77 GLY CA 28.96 28.77 42.33 15.00
77 GLY C 30.07 29.15 41.36 15.00
77 GLY 0 30.68 30.20 41.51 15.00
78 LEU N 30.32 28.33 40.35 15.00
78 LEU CA 31.38 28.66 39.42 15.00
78 LEU CB 31.85 27.45 38.61 15.00
78 LEU CG 33.00 27.84 37.67 15.00
78 LEU CDl 34.30 27.78 38.44 15.00
78 LEU CD2 33.07 26.95 36.46 15.00
78 LEU C 30.86 29.69 38.47 15.00
78 LEU 0 30.29 29.33 37.44 15.00 79 LYS N 31.00 30.96 38.81 15.00
79 LYS CA 30.57 32.02 37.93 15.00
79 LYS CB 30.77 33.37 38.60 15.00
79 LYS CG 29.87 34.48 38.12 15.00
79 LYS CD 28.54 34.41 38.82 15.00
79 LYS CE 27.78 35.74 38.71 15.00
79 LYS NZ 28.35 36.82 39.58 15.00
79 LYS C 31.52 31.90 36.75 15.00
79 LYS O 32.58 32.52 36.75 15.00
80 VAL N 31.18 31.04 35.78 15.00
80 VAL CA 32.02 30.81 34.61 15.00
80 VAL CB 31.23 30.06 33.49 15.00
80 VAL CGI 30.41 31.00 32.66 15.00
80 VAL CG2 32.14 29.23 32.63 15.00
80 VAL C 32.57 32.17 34.16 15.00
80 VAL O 31.87 33.19 34.25 15.00
81 PRO N 33.84 32.21 33.74 15.00
81 PRO CD 34.68 31.03 33.50 15.00
81 PRO CA 34.53 33.42 33.29 15.00
81 PRO CB 35.96 32.92 33.09 15.00
81 PRO CG 35.73 31.56 32.54 15.00
81 PRO C 34.00 34.12 32.03 15.00
81 PRO O 32.79 34.26 31.82 15.00
82 LEU N 34.96 34.60 31.23 15.00
82 LEU CA 34.73 35.30 29.97 15.00
82 LEU CB 34.38 36.77 30.24 15.00
82 LEU CG 34.75 37.85 29.22 15.00
82 LEU CDl 33.96 37.69 27.91 15.00
82 LEU CD2 34.52 39.22 29.85 15.00
82 LEU C 36.07 35.17 29.23 15.00
82 LEU O 37.05 34.72 29.83 15.00
83 SER N 36.12 35.54 27.95 15.00
83 SER CA 37.35 35.45 27.17 15.00
83 SER CB 37.17 36.03 25.77 15.00
83 SER OG 36.98 34.98 24.81 15.00
83 SER C 38.68 35.92 27.78 15.00
83 SER O 38.78 37.02 28.37 15.00 84 HIS N 39.67 35.05 27.61 15.00
84 HIS CA 41.04 35.22 28.07 15.00
84 HIS CB 41.09 35.25 29.61 15.00
84 HIS CG 42.39 35.75 30.18 15.00
84 HIS CD2 42.72 36.94 '30.75 15.00
84 HIS NDl 43.55 34.99 30.19 15.00
84 HIS CEl 44.52 35.69 30.75 15.00
84 HIS NE2 44.05 36.88 31.09 15.00
84 HIS C 41.90 34.05 27.56 15.00
84 HIS 0 42.85 33.65 28.23 15.00
85 SER N 41.54 33.48 26.41 15.00
85 SER CA 42.33 32.37 25.84 15.00
85 SER CB 42.43 31.19 26.83 15.00
85 SER OG 43.22 30.13 26.30 15.00
85 SER C 41.91 31.83 24.46 15.00
85 SER 0 40.73 31.77 24.12 15.00
86 ARG N 42.92 31.36 23.73 15.00
86 ARG CA 42.74 30.80 22.39 15.00
86 ARG CB 42.47 31.93 21.36 15.00
86 ARG CG 41.83 31.46 20.04 15.00
86 ARG CD 40.35 31.05 20.25 15.00
86 ARG NE 39.78 30.29 19.13 15.00
86 ARG CZ 39.86 30.63 17.84 15.00
86 ARG NHl 40.50 31.73 17.46 15.00
86 ARG NH2 39.30 29.84 16.91 15.00
86 ARG C 44.03 30.02 22.03 15.00
86 ARG 0 44.51 30.07 20.90 15.00
87 SER N 44.60 29.36 23.03 15.00
87 SER CA 45.80 28.55 22.84 15.00
87 SER CB 45.48 27.28 22.04 15.00
87 SER OG 44.46 26.51 22.68 15.00
87 SER C 47.06 29.24 22.27 15.00
87 SER 0 47.65 28.76 21.30 15.00
88 ASN N 47.47 30.35 22.88 15.00
88 ASN CA 48.68 31.07 22.43 15.00
88 ASN CB 48.65 31.36 20.90 15.00
88 ASN CG 47.56 32.38 20.49 15.00 88 ASN ODl 46.95 33.06 21.33 15.00
88 ASN ND2 47, .33 32 .49 19 .18 15 .00
88 ASN C 48, .96 32 .35 23 .21 15. .00
88 ASN 0 49 .83 33 .15 22 .85 15 .00
89 ASP N 48 .24 32 .53 24 .32 15. .00
89 ASP CA 48, .40 33 .72 25 .14 15. .00
89 ASP CB 47, .28 33 .81 26 .18 15. .00
89 ASP CG 46, .34 34, .97 25 .92 15, .00
89 ASP ODl 46, .63 36 .09 .26 .41 15. .00
89 ASP OD2 45, .31 34 .76 25 .24 15 .00
89 ASP C 49, .75 33. .75 25 .85 15, .00
89 ASP 0 50, .62 32, .93 25 .58 15, .00
90 THR N 49 .87 34 .71 26 .76 15 .00
90 THR CA 51, .07 34 .91 27 .57 15, .00
90 THR CB 51. .26 36 .42 27 .89 15, .00
90 THR OGl 50, .27 36, .83 28, .83 15. .00
90 THR CG2 51. .10 37 .26 26, .62 15, .00
90 THR C 50, .84 34, .14 28, .87 15, .00
90 THR 0 51, .75 33, .52 29, .41 15. .00
91 LEU N 49. ,61 34, .23 29, .39 15. .00
91 LEU CA 49. .23 33. .52 30. ,60 15. ,00
91 LEU CB 48. .31 34, .38 31. .47 15. .00
91 LEU CG 48. .82 34, .78 32, .87 15. ,00
91 LEU CDl 50. .36 34. .84 ■ 32. .93 15. ,00
91 LEU CD2 48. ,21 36. .13 33. ,25 15. .00
91 LEU C 48, .48 32. .31 30. .08 15. .00
91 LEU 0 47. .26 32. .29 30. .04 15. .00
92 TYR N 49. ,24 31. .31 29. .65 15. .00
92 TYR CA 48. .71 30. .09 29. ,06 15. ,00
92 TYR CB 48. ,45 30. ,39 27. ,59 15. ,00
92 TYR CG 48. ,08 29. .20 26. .76 15. ,00
92 TYR CDl 49. ,05 28, .36 26. ,26 15. ,00
92 TYR CEl 48. .73 27. ,27 25. ,45 15. ,00
92 TYR CD2 46. ,74 28. .95 26. ,43 15. 00
92 TYR CE2 46. ,40 27. .87 25. .63 15. ,00
92 TYR CZ 47. ,40 27. ,03 25. ,14 15. 00
92 TYR OH 47. ,09 25. .97 24. ,30 15. 00 92 TYR C 49.84 29.07 29.22 15.00
92 TYR 0 51.00 29.41 29.01 15.00
93 ILE N 49.53 27.82 29.57 15.00
93 ILE CA 50.61 26.86 29.78 15.00
93 ILE CB 50.50 26.15 31.14 15.00
93 ILE CG2 49.45 25.09 31.09 15.00
93 ILE CGI 51.80 25.44 31.46 15.00
93 ILE CDl 51.84 24.86 32.84 15.00
93 ILE C 50.90 25.82 28.72 15.00
93 ILE 0 50.02 25.09 28.30 15.00
94 PRO N 52.12 25.85 28.18 15.00
94 PRO CD 52.88 27.11 28.07 15.00
94 PRO CA 52.60 24.92 27.16 15.00
94 PRO CB 53.31 25.83 '26.15 15.00
94 PRO CG 52.95 27.26 26.58 15.00
94 PRO C 53.61 24.06 27.92 15.00
94 PRO 0 54.61 24.57 28.43 15.00
95 GLU N 53.33 22.78 28.07 15.00
95 GLU CA 54.25 21.92 28.80 15.00
95 GLU CB 53.53 20.64 29.23 15.00
95 GLU CG 52.19 20.88 29.95 15.00
95 GLU CD 52.28 21.71 31.25 15.00
95 GLU OEl 51.22 22.21 31.70 15.00
95 GLU OE2 53.39 21.86 31.83 15.00
95 GLU C 55.46 21.61 27.91 15.00
95 GLU 0 55.30 21.42 26.70 15.00
96 TRP N 56.66 21.58 28.49 15.00
96 TRP CA 57.88 21.32 27.71 15.00
96 TRP CB 59.09 21.98 28.38 15.00
96 TRP CG 59.20 21.70 29.82 15.00
96 TRP CD2 59.29 22.67 30.87 15.00
96 TRP CE2 59.44 21.96 32.09 15.00
96 TRP CE3 59.26 24.07 30.89 15.00
96 TRP CDl 59.29 20.48 30.43 15.00
96 TRP NEl 59.44 20.62 31.79 15.00
96 TRP CZ2 59.56 22.61 33.32 15.00
96 TRP CZ3 59.38 24.71 32.12 15.00 96 TRP CH2 59.53 23.98 33.31 15.00
96 TRP C 58.18 19.86 27.43 15.00
96 TRP O 57.89 19.00 .28.24 15-00
97 GLU N 58.87 19.64 26.30 15.00
97 GLU CA 59.28 18.33 25.77 15.00
97 GLU CB 60.24 18.53 24.59 15.00
97 GLU CG 59.86 19.64 23.61 15.00
97 GLU CD 61.09 20.40 23.09 15.00
97 GLU OEl 61.77 19.91 22.15 15.00
97 GLU OE2 61.36 21.49 23.64 15.00
97 GLU C 59.89 17.31 26.75 15.00
97 GLU O 61.02 16.83 26.56 15.00
98 GLY N 59.08 16.92 27.73 15.00
98 GLY CA 59.49 15.97 28.75 15.00
98 GLY C 58.65 14.71 28.66 15.00
98 GLY O 58.23 14.30 27.58 15.00
99 ARG N 58.27 14.18 29.81 15.00
99 ARG CA 57.54 12.92 •29.84 15.00
99 ARG CB 58.60 11.81 29.82 15.00
99 ARG CG 58.16 10.40 29.50 15.00
99 ARG CD 59.36 9.55 29.09 15.00
99 ARG NE 59.78 9.84 27.71 15.00
99 ARG CZ 60.90 10.48 27.36 15.00
99 ARG NHl 61.75 10.93 28.28 15.00
99 ARG NH2 61.16 10.69 26.07 15.00
99 ARG C 56.79 12.95 31.18 15.00
99 ARG O 57.15 13.75 32.04 15.00
100 ALA N 55.77 12.11 31.36 15.00
100 ALA CA 55.04 12.13 32.64 15.00
100 ALA CB 53.72 12.88 32.49 15.00
100 ALA C 54.81 10.77 33.28 15.00
100 ALA O 54.45 9.79 32.61 15.00
101 PRO N 54.99 10.69 •34.61 15.00
101 PRO CD 55.38 11.79 35.49 15.00
101 PRO CA 54.80 9.46 35.38 15.00
101 PRO CB 55.16 9.89 36.80 15.00
101 PRO CG 54.85 11.34 36.81 15.00 101 PRO C 53.40 8.87 35.31 15.00
101 PRO 0 52.44 9.56 34.97 15.00
102 ASP N 53.30 7.59 35.64 15.00
102 ASP CA 52.03 6.86 35.66 15.00
102 ASP CB 52.30 5.36 35.82 15.00
102 ASP CG 52.72 4.70 34.51 15.00
102 ASP ODl 53.12 5.39 33.54 15.00
102 ASP OD2 52.63 3.45 '34.45 15.00
102 ASP C 51.16 7.36 36.83 15.00
102 ASP 0 50.09 7.92 36.62 15.00
103 SER N 51.62 7.10 38.05 15.00
103 SER CA 50.93 7.51 39.27 15.00
103 SER CB 51.43 6.64 40.44 15.00
103 SER OG 50.89 7.02 41.70 15.00
103 SER C 51.28 8.99 39.51 15.00
103 SER O 51.87 9.62 38.63 15.00
104 VAL N 50.86 9.54 40.66 15.00
104 VAL CA 51.13 10.93 41.06 15.00
104 VAL CB 51.10 11.88 39.85 15.00
104 VAL CGI 49.67 12.03 39.37 15.00
104 VAL CG2 51.76 13.20 40.18 15.00
104 VAL C 50.14 11.47 42.12 15.00
104 VAL O 49.77 12.63 '42.11 15.00
105 ASP N 49.76 10.64 43.06 15.00
105 ASP CA 48.82 11.06 44.09 15.00
105 ASP CB 48.39 9.82 44.87 15.00
105 ASP CG 47.34 10.10 45.93 15.00
105 ASP ODl 47.33 11.19 46.55 15.00
105 ASP OD2 46.53 9.18 46.17 15.00
105 ASP C 49.54 12.05 44.99 15.00
105 ASP O 50.36 11.64 45.82 15.00
106 TYR N 49.23 13.34 44.89 15.00
106 TYR CA 49.88 14.34 45.73 15.00
106 TYR CB 49.55 15.74 45.28 15.00
106 TYR CG 50.43 16.20 44.17 15.00
106 TYR CDl 51.75 16.55 44.42 15.00
106 TYR CEl 52.58 16.95 43.38 15.00 106 TYR CD2 49.96 16.25 42.87 15.00
106 TYR CE2 50.78 16.65 41.83 15.00
106 TYR CZ 52.09 16.99 42.09 15.00
106 TYR OH 52.89 17.37 41.04 15.00
106 TYR C 49.66 14.23 47.23 15.00
106 TYR 0 50.35 14.88 48.03 15.00
107 ARG N 48.66 13.45 47.63 15.00
107 ARG CA 48.40 13.29 49.05 15.00
107 ARG CB 47.01 12.70 49.26 15.00
107 ARG CG 45.92 13.66 48.83 15.00
107 ARG CD 44.62 12.97 48.69 15.00
107 ARG NE 44.58 12.10 ■47.52 15.00
107 ARG CZ 43.98 10.92 47.51 15.00
107 ARG NHl 43.38 10.47 48.61 15.00
107 ARG NH2 43.90 10.22 46.38 15.00
107 ARG C 49.50 12.42 49.66 15.00
107 ARG 0 49.93 12.65 50.79 15.00
108 LYS N 49.99 11.46 48.88 15.00
108 LYS CA 51.09 10.60 49.32 15.00
108 LYS CB 51.48 9.59 48.24 15.00
108 LYS CG 50.50 8.45 48.02 15.00
108 LYS CD 50.82 7.25 48.91 15.00
108 LYS CE 49.90 6.05 48.60 15.00
108 LYS NZ 49.85 5.66 47.15 15.00
108 LYS C 52.26 11.56 49.53 15.00
108 LYS 0 52.99 11.46 50.51 15.00
109 LYS N 52.37 12.52 ■48.61 15.00
109 LYS CA 53.43 13.53 48.62 15.00
109 LYS CB 53.61 14.12 47.22 15.00
109 LYS CG 53.60 13.06 46.12 15.00
109 LYS CD 53.94 13.64 44.75 15.00
109 LYS CE 55.40 14.03 44.66 15.00
109 LYS NZ 56.29 12.84 44.80 15.00
109 LYS C 53.17 14.65 49.63 15.00
109 LYS 0 53.76 15.72 49.54 15.00
110 GLY N 52.23 14.41 50.54 15.00
110 GLY CA 51.88 15.35 51.59 15.00 110 GLY C 51.79 16.81 .51.26 15.00
110 GLY O 52.16 17.64 52.07 15.00
111 TYR N 51.28 17.15 50.08 15.00
111 TYR CA 51.13 18.55 49.66 15.00
111 TYR CB 51.43 18.71 48.15 15.00
111 TYR CG 52.89 18.57 47.73 15.00
111 TYR CDl 53.87 18.20 48.65 15.00
111 TYR CEl 55.20 18.05 48.27 15.00
111 TYR CD2 53.29 18.80 46.41 15.00
111 TYR CE2 54.62 18.66 46.02 15.00
111 TYR CZ 55.57 18.27 46.97 15.00
111 TYR OH 56.89 18.08 46.63 15.00
111 TYR C 49.73 19.14 49.94 15.00
111 TYR O 49.54 20.36 49.92 15.00
112 VAL N 48.77 18.28 50.26 15.00
112 VAL CA 47.38 18.71 .50.45 15.00
112 VAL CB 46.46 17.87 49.56 15.00
112 VAL CGI 45.13 18.59 49.37 15.00
112 VAL CG2 47.13 17.52 48.23 15.00
112 VAL C 46.81 18.70 51.87 15.00
112 VAL O 46.97 17.73 52.63 15.00
113 THR N 46.10 19.78 52.20 15.00
113 THR CA 45.47 19.93 53.51 15.00
113 THR CB 45.05 21.38 53.76 15.00
113 THR OGl 44.71 22.00 52.52 15.00
113 THR CG2 46.15 22.14 54.46 15.00
113 THR C 44.20 19.09 53.59 15.00
113 THR O 43.77 18.50 52.60 15.00
114 PRO N 43.60 19.01 54.79 15.00
114 PRO CD 44.13 19.46 56.09 15.00
114 PRO CA 42.37 18.25 .54.99 15.00
114 PRO CB 42.09 18.45 56.47 15.00
114 PRO CG 43.42 18.55 57.06 15.00
114 PRO C 41.24 18.82 54.13 15.00
114 PRO O 41.36 19.94 53.62 15.00
115 VAL N 40.16 18.05 53.97 15.00
115 VAL CA 39.00 18.50 53.19 15.00 115 VAL CB 38.13 17.31 52.69 15.00
115 VAL CGI 36.92 17.83 51.93 15.00
115 VAL CG2 38.93 16.41 51.77 15.00
115 VAL C 38.18 19.38 54.11 15.00
115 VAL 0 37.77 18.97 55.19 15.00
116 LYS N 38.01 20.63 •53.70 15.00
116 LYS CA 37.24 21.56 54.49 15.00
116 LYS CB 37.79 22.98 54.34 15.00
116 LYS CG 39.28 23.07 54.08 15.00
116 LYS CD 39.54 23.07 52.58 15.00
116 LYS CE 41.03 23.20 52.27 15.00
116 LYS NZ 41.56 24.48 52.82 15.00
116 LYS C 35.81 21.46 53.97 15.00
116 LYS 0 35.55 20.71 53.03 15.00
117 ASN N 34.89 22.21 54.57 15.00
117 ASN CA 33.50 22.18 54.13 15.00
117 ASN CB 32.62 21.57 55.21 15.00
117 ASN CG 31.16 21.70 54.88 15.00
117 ASN ODl 30.61 20.93 54.10 15.00
117 ASN ND2 30.52 22.71 55.46 15.00
117 ASN C 33.02 23.59 ■53.81 15.00
117 ASN 0 32.92 24.43 54.70 15.00
118 GLN N 32.65 23.81 52.55 15.00
118 GLN CA 32.21 25.12 52.08 15.00
118 GLN CB 32.17 25.14 50.55 15.00
118 GLN CG 30.98 24.41 49.94 15.00
118 GLN CD 30.83 24.71 48.46 15.00
118 GLN OEl 31.38 24.01 47.61 15.00
118 GLN NE2 30.11 25.77 48.14 15.00
118 GLN C 30.86 25.59 52.59 15.00
118 GLN 0 30.32 26.57 52.09 15.00
119 GLY N 30.32 24.91 53.58 15.00
119 GLY CA 29.02 25.31 54.10 15.00
119 GLY C 28.03 25.31 52.96 15.00
119 GLY 0 28.21 24.61 51.97 15.00
120 GLN N 27.01 26.16 •53.06 15.00
120 GLN CA 25.99 26.23 52.04 15.00 120 GLN CB 24.60 25.99 52.65 15.00
120 GLN CG 24.28 26.85 53.89 15.00
120 GLN CD 25.10 26.50 55.13 15.00
120 GLN OEl 26.03 27.22 55.52 15.00
120 GLN NE2 24.75 25.38 55.76 15.00
120 GLN C 26.10 27.61 51.41 15.00
120 GLN 0 25.12 28.34 51.29 15.00
121 CYS N 27.33 27.97 51.04 15.00
121 CYS CA 27.63 29.26 50.43 15.00
121 CYS C 28.38 29.01 49.13 15.00
121 CYS 0 28.91 27.93 ■48.92 15.00
121 CYS CB 28.43 30.13 51.42 15.00
121 CYS SG 29.68 31.27 50.76 15.00
122 GLY N 28.41 29.98 48.24 15.00
122 GLY CA 29.08 29.76 46.98 15.00
122 GLY C 30.58 29.94 47.04 15.00
122 GLY 0 31.13 30.69 46.24 15.00
123 SER N 31.24 29.25 47.96 15.00
123 SER CA 32.69 29.37 48.08 15.00
123 SER CB 33.10 29.19 49.53 15.00
123 SER OG 32.79 30.33 50.30 15.00
123 SER C 33.44 28.36 47.23 15.00
123 SER 0 34.63 28.16 47.44 15.00
124 CYS N 32.79 27.73 46.27 15.00
124 CYS CA 33.49 26.74 45.45 15.00
124 CYS CB 32.55 26.04 •44.46 15.00
124 CYS SG 32.47 26.70 42.78 15.00
124 CYS C 34.71 27.36 44.77 15.00
124 CYS 0 35.80 26.79 44.79 15.00
125 TRP N 34.54 28.58 44.25 15.00
125 TRP CA 35.62 29.31 43.58 15.00
125 TRP CB 35.11 30.63 42.97 15.00
125 TRP CG 34.60 31.64 43.98 15.00
125 TRP CD2 35.30 32.77 44.49 15.00
125 TRP CE2 34.48 33.37 45.47 15.00
125 TRP CE3 36.56 33.34 44.23 15.00
125 TRP CDl 33.40 31.61 44.63 15.00 125 TRP NEl 33.33 32.64 45.53 15.00
125 TRP CZ2 34.87 34.51 46.18 15.00
125 TRP CZ3 36.95 34.47 44.94 15.00
125 TRP CH2 36.10 35.04 45.90 15.00
125 TRP C 36.72 29.59 44.60 15.00
125 TRP 0 37.89 29.34 44.34 15.00
126 ALA N 36.33 30.05 45.79 15.00
126 ALA CA 37.30 30.34 46.83 15.00
126 ALA CB 36.68 31.21 47.90 15.00
126 ALA C 37.90 29.09 47.45 15.00
126 ALA 0 38.80 29.19 48.28 15.00
127 PHE N 37.39 27.92 47.12 15.00
127 PHE CA 37.98 26.72 47.69 15.00
127 PHE CB 36.92 25.73 48.16 15.00
127 PHE CG 36.48 25.95 .49.59 15.00
127 PHE CDl 35.54 26.93 49.90 15.00
127 PHE CD2 37.03 25.21 50.62 15.00
127 PHE CEl 35.17 27.16 51.21 15.00
127 PHE CE2 36.66 25.45 51.95 15.00
127 PHE CZ 35.72 26.42 52.24 15.00
127 PHE C 38.98 26.09 46.74 15.00
127 PHE 0 40.05 25.67 47.16 15.00
128 SER N 38.66 26.08 45.46 15.00
128 SER CA 39.56 25.54 44.45 15.00
128 SER CB 38.91 25.64 43.07 15.00
128 SER OG 39.60 24.88 42.09 15.00
128 SER C 40.85 26.35 44.50 15.00
128 SER 0 41.95 25.80 44.63 15.00
129 SER N 40.72 27.68 44.44 15.00
129 SER CA 41.86 28.57 .44.47 15.00
129 SER CB 41.40 30.01 44.43 15.00
129 SER OG 40.65 30.23 43.25 15.00
129 SER C 42.69 28.33 45.69 15.00
129 SER 0 43.87 28.02 45.59 15.00
130 VAL N 42.07 28.41 46.85 15.00
130 VAL CA 42.79 28.18 48.10 15.00
130 VAL CB 41.81 28.14 49.28 15.00 130 VAL CGI 42.27 27.18 50.35 15.00
130 VAL CG2 41.70 29.52 49.85 15.00
130 VAL C 43.58 26.88 48.04 15.00
130 VAL 0 44.65 26.79 48.62 15.00
131 GLY N 43.08 25.91 '47.29 15.00
131 GLY CA 43.74 24.61 47.18 15.00
131 GLY C 45.07 24.66 46.45 15.00
131 GLY 0 46.06 24.12 46.94 15.00
132 ALA N 45.07 25.26 45.27 15.00
132 ALA CA 46.27 25.39 44.47 15.00
132 ALA CB 45.92 26.00 43.15 15.00
132 ALA C 47.30 26.25 45.21 15.00
132 ALA 0 48.51 26.01 45.13 15.00
133 LEU N 46.82 27.26 45.93 15.00
133 LEU CA 47.70 28.13 46.69 15.00
133 LEU CB 46.92 29.27 47.32 15.00
133 LEU CG 46.66 30.36 46.31 15.00
133 LEU CDl 45.87 31.46 46.98 15.00
133 LEU CD2 47.97 30.90 45.76 15.00
133 LEU C 48.46 27.36 47.76 15.00
133 LEU 0 49.65 27.58 47.96 15.00
134 GLU N 47.77 26.45 48.43 15.00
134 GLU CA 48.40 25.64 49.46 15.00
134 GLU CB 47.35 24.78 50.17 15.00
134 GLU CG 46.35 25.63 50.99 15.00
134 GLU CD 45.02 24.94 51.29 15.00
134 GLU OEl 44.30 25.44 52.17 15.00
134 GLU OE2 44.69 23.92 50.65 15.00
134 GLU C 49.52 24.78 48.86 15.00
134 GLU 0 50.67 24.91 49.26 15.00
135 GLY N 49.18 24.01 47.83 15.00
135 GLY CA 50.13 23.13 47.15 15.00
135 GLY C 51.43 23.81 46.79 15.00
135 GLY 0 52.51 23.19 46.84 15.00
136 GLN N 51.33 25.08 46.41 15.00
136 GLN CA 52.49 25.88 46.06 15.00
136 GLN CB 52.04 27.14 45.34 15.00 136 GLN CG 51.39 26.82 44.03 15.00
136 GLN CD 52.23 25.85 43.24 15.00
136 GLN OEl 52.02 24.64 43.33 15.00
136 GLN NE2 53.23 26.36 42.53 15.00
136 GLN C 53.23 26.24 47.34 15.00
136 GLN 0 54.44 26.02 47.45 15.00
137 LEU N 52.50 26.74 48.34 15.00
137 LEU CA 53.10 27.10 49.62 15.00
137 LEU CB 52.02 27.35 •50.67 15.00
137 LEU CG 52.49 27.78 52.05 15.00
137 LEU CDl 53.16 29.11 51.94 15.00
137 LEU CD2 51.32 27.87 52.99 15.00
137 LEU C 53.98 25.94 50.08 15.00
137 LEU 0 55.10 26.14 50.55 15.00
138 LYS N 53.49 24.73 49.87 15.00
138 LYS CA 54.23 23.54 50.25 15.00
138 LYS CB 53.37 22.30 50.06 15.00
138 LYS CG 54.09 21.00 50.39 15.00
138 LYS CD 54.37 20.87 51.88 15.00
138 LYS CE 54.98 19.52 52.20 15.00
138 LYS NZ 55.22 19.34 53.66 15.00
138 LYS C 55.48 23.42 49.39 15.00
138 LYS 0 56.60 23.42 49.90 15.00
139 LYS N 55.27 23.37 •48.08 15.00
139 LYS CA 56.35 23.22 47.12 15.00
139 LYS CB 55.76 22.98 45.72 15.00
139 LYS CG 56.78 22.68 44.61 15.00
139 LYS CD 56.11 22.53 43.25 15.00
139 LYS CE 57.10 22.11 42.18 15.00
139 LYS NZ 56.47 21.93 40.84 15.00
139 LYS C 57.36 24.36 47.10 15.00
139 LYS 0 58.36 24.27 46.40 15.00
140 LYS N 57.13 25.43 47.87 15.00
140 LYS CA 58.07 26.55 47.86 15.00
140 LYS CB 57.47 27.72 47.08 15.00
140 LYS CG 58.47 28.78 46.62 15.00
140 LYS CD 57.75 29.84 45.80 15.00 140 LYS CE 58.71 30.80 45.06 15.00
140 LYS NZ 59.20 30.29 43.72 15.00
140 LYS C 58.47 27.02 49.26 15.00
140 LYS 0 59.15 28.04 49.40 15.00
141 THR N 58.06 26.28 50.28 15.00 141 THR CA 58.40 26.63 51.67 15.00 141 THR CB 57.41 27.66 52.27 15.00 141 THR OGl 57.48 28.88 51.52 15.00 141 THR CG2 57.73 27.97 53.72 15.00 141 THR C 58.51 25.38 52.55 15.00
141 THR 0 58.92 25.45 53.71 15.00
142 GLY N 58.16 24.22 51.99 15.00 142 GLY CA 58.27 22.97 52.72 15.00 142 GLY C 57.25 22.78 53.83 15.00
142 GLY 0 57.03 21.67 54.29 15.00
143 LYS N 56.72 23.89 54.33 15.00 143 LYS CA 55.69 23.84 55.34 15.00 143 LYS CB 55.76 25.09 56.22 15.00 143 LYS CG 56.85 25.06 57.26 15.00 143 LYS CD 56.33 24.57 58.61 15.00 143 LYS CE 55.35 25.58 59.22 15.00 143 LYS NZ 54.85 25.16 60.56 15.00 143 LYS C 54.37 23.83 54.58 15.00
143 LYS 0 54.32 24.20 53.40 15.00
144 LEU N 53.31 23.39 55.23 15.00 144 LEU CA 52.00 23.37 54.60 15.00 144 LEU CB 51.70 21.99 54.02 15.00 144 LEU CG 50.28 21.43 54.19 15.00 144 LEU CDl 49.90 20.63 52.95 15.00 144 LEU CD2 50.19 20.58 55.46 15.00 144 LEU C 50.97 23.72 55.65 15.00
144 LEU 0 50.97 23.14 56.74 15.00
145 LEU N 50.16 24.72 55.36 15.00 145 LEU CA 49.12 25.12 56.28 15.00 145 LEU CB 49.59 26.22 57.23 15.00 145 LEU CG 50.20 27.54 56.77 15.00 145 LEU CDl 50.04 28.57 57.88 15.00 145 LEU CD2 51.68 27.36 56.43 15.00
145 LEU C 47.89 25.54 55.49 15.00
145 LEU O 47.95 25.73 •54.27 15.00
146 ASN N 46.77 25.64 56.20 15.00
146 ASN CA 45.51 26.02 55.57 15.00
146 ASN CB 44.33 25.53 56.41 15.00
146 ASN CG 44.65 24.27 57.19 15.00
146 ASN ODl 45.62 23.58 56.90 15.00
146 ASN ND2 43.84 23.99 58.20 15.00
146 ASN C 45.40 27.52 55.41 15.00
146 ASN O 45.53 28.27 56.39 15.00
147 LEU N 45.16 27.96 54.18 15.00
147 LEU CA 44.99 29.36 53.84 15.00
147 LEU CB 45.51 29.62 52.43 15.00
147 LEU CG 46.91 29.08 52.19 15.00
147 LEU CDl 47.34 29.38 50.77 15.00
147 LEU CD2 47.85 29.70 53.18 15.00
147 LEU C 43.49 29.62 53.91 15.00
147 LEU O 42.71 28.70 53.68 15.00
148 SER N 43.10 30.85 54.20 15.00
148 SER CA 41.68 31.21 54.31 15.00
148 SER CB 41.52 32.50 55.12 15.00
148 SER OG 40.20 32.98 55.01 15.00
148 SER C 40.90 31.36 53.00 15.00
148 SER O 41.20 32.22 52.18 15.00
149 PRO N 39.82 30.57 52.85 15.00
149 PRO CD 39.42 29.53 53.81 15.00
149 PRO CA 38.92 30.56 51.69 15.00
149 PRO CB 38.09 29.30 51.91 15.00
149 PRO CG 38.90 28.48 52.90 15.00
149 PRO C 38.04 31.80 51.73 15.00
149 PRO O 37.68 32.37 50.70 15.00
150 GLN N 37.72 32.23 52.94 15.00
150 GLN CA 36.88 33.41 53.09 15.00
150 GLN CB 36.42 33.61 54.53 15.00
150 GLN CG 35.40 34.72 54.62 15.00
150 GLN CD 34.35 34.51 55.69 15.00 150 GLN OEl 33,.83 33..41 55,.87 15,.00
150 GLN NE2 34. .01 35. .59 56, .37 15, .00
150 GLN C 37. .59 34. .65 52, .55 15 .00
150 GLN 0 37. .05 35. .31 51, .67 15, .00
151 ASN N 38, .79 34, .94 53, .06 15, .00
151 ASN CA 39, .60 36. .10 52, .65 15, .00
151 ASN CB 41, .10 35. .74 '52. .72 15, .00
151 ASN CG 42, .01 36. .96 52, .57 15, .00
151 ASN ODl 42, .13 37. .55 51. .50 15. .00
151 ASN ND2 42, .69 37. .31 53, .65 15, .00
151 ASN C 39, .26 36. .44 51. .22 15. .00
151 ASN 0 38, .89 37, .57 50, .91 15, .00
152 LEU N 39. .30 35, .42 50, .38 15. .00
152 LEU CA 38. .97 35, .57 48, .99 15, .00
152 LEU CB 39, .13 34, .25 48, .23 15. .00
152 LEU CG 40, .51 33, .63 48, .09 15, .00
152 LEU CDl 41, .50 34. .67 47, .60 15. .00
152 LEU CD2 40. .95 33. .07 49. .41 15, .00
152 LEU C 37, .53 36. .04 48. .93 15. .00
152 LEU 0 37, .29 37. .20 48. .60 15. .00
153 VAL N 36, .59 35. .19 49. .34 15. ,00
153 VAL CA 35. .16 35. .50 '49. .32 15, .00
153 VAL CB 34, .35 34, .61 50, .31 15. .00
153 VAL CGI 32. .85 34. .96 50. .23 15. ,00
153 VAL CG2 34. .56 33, .13 50, .03 15. .00
153 VAL C 34, .78 36. .95 49. .62 15, ,00
153 VAL 0 34, .07 37. .57 48, .82 15. .00
154 ASP N 35. .21 37. .46 50. .77 15. ,00
154 ASP CA 34, .89 38. .84 51. .17 15. ,00
154 ASP CB 35. .15 39. .06 52. .67 15. ,00
154 ASP CG 34. .67 37. ,94 53. ,54 15. .00
154 ASP ODl 34. .06 36. .98 53. .01 15. .00
154 ASP OD2 34. .91 38. .00 54. ,77 15. .00
154 ASP C 35. .68 39. .91 50, .44 15. ,00
154 ASP 0 35, .16 41. .00 50. ,20 15. ,00
155 CYS N 36. .95 39. .64 50, ,15 15. .00
155 CYS CA 37, .78 40. .64 '49. .51 15. ,00 155 CYS C 37.79 40.73 47.99 15.00
155 CYS 0 37.81 41.83 47.45 15.00
155 CYS CB 39.17 40.56 50.08 15.00
155 CYS SG 39.15 40.52 51.90 15.00
156 VAL N 37.82 39.59 47.30 15.00
156 VAL CA 37.83 39.60 45.84 15.00
156 VAL CB 37.88 38.17 45.26 15.00
156 VAL CGI 37.89 38.22 43.75 15.00
156 VAL CG2 39.11 37.48 45.75 15.00
156 VAL C 36.59 40.33 45.30 15.00
156 VAL 0 35.64 39.70 .44.83 15.00
157 SER N 36.64 41.65 45.31 15.00
157 SER CA 35.53 42.48 44.85 15.00
157 SER CB 35.92 43.96 44.90 15.00
157 SER OG 37.07 44.17 45.70 15.00
157 SER C 35.10 42.14 43.45 15.00
157 SER 0 34.07 42.63 42.99 15.00
158 GLU N 35.90 41.36 42.74 15.00
158 GLU CA 35.56 40.99 41.37 15.00
158 GLU CB 36.83 40.69 40.56 15.00
158 GLU CG 37.78 41.88 40.35 15.00
158 GLU CD 39.15 41.67 41.00 15.00
158 GLU OEl 39.62 40.51 41.07 15.00
158 GLU OE2 39.76 42.68 41.46 15.00
158 GLU C 34.63 39.77 41.34 15.00
158 GLU 0 34.06 39.43 .40.30 15.00
159 ASN N 34.46 39.12 42.49 15.00
159 ASN CA 33.62 37.93 42.57 15.00
159 ASN CB 34.47 36.70 42.94 15.00
159 ASN CG 35.27 36.15 41.75 15.00
159 ASN ODl 36.23 35.41 41.95 15.00
159 ASN ND2 34.85 36.48 40.53 15.00
159 ASN C 32.45 38.09 43.53 15.00
159 ASN 0 32.57 38.70 44.59 15.00
160 ASP N 31.32 37.49 43.17 15.00
160 ASP CA 30.10 37.60 43.97 15.00
160 ASP CB 28.91 37.01 43.19 15.00 160 ASP CG 27.72 37.96 ■43.13 15.00
160 ASP ODl 27.04 37.99 42.07 15.00
160 ASP OD2 27.47 38.68 44.12 15.00
160 ASP C 30.16 36.96 45.35 15.00
160 ASP 0 29.12 36.80 46.00 15.00
161 GLY N 31.35 36.59 45.80 15.00
161 GLY CA 31.46 35.98 47.11 15.00
161 GLY C 30.58 34.75 47.20 15.00
161 GLY 0 30.57 33.93 46.28 15.00
162 CYS N 29.77 34.67 48.25 15.00
162 CYS CA 28.88 33.53 48.42 15.00
162 CYS C 27.91 33.35 47.27 15.00
162 CYS 0 27.27 32.31 47.16 15.00
162 CYS CB 28.11 33.63 49.74 15.00
162 CYS SG 29.06 33.14 51.22 15.00
163 GLY N 27.80 34.35 •46.40 15.00
163 GLY CA 26.89 34.24 45.29 15.00
163 GLY C 27.55 33.80 44.00 15.00
163 GLY 0 26.94 33.84 42.93 15.00
164 GLY N 28.81 33.41 44.09 15.00
164 GLY CA 29.51 32.96 42.90 15.00
164 GLY C 30.80 33.70 42.64 15.00
164 GLY 0 31.08 34.75 43.23 15.00
165 GLY N 31.60 33.14 41.73 15.00
165 GLY CA 32.87 33.73 41.37 15.00
165 GLY C 33.69 32.80 40.50 15.00
165 GLY 0 33.15 31.84 39.94 15.00
166 TYR N 34.99 33.03 40.42 15.00
166 TYR CA 35.87 32.22 39.60 15.00
166 TYR CB 35.91 32.77 38.19 15.00
166 TYR CG 36.63 31.87 '37.25 15.00
166 TYR CDl 37.97 32.08 36.95 15.00
166 TYR CEl 38.64 31.27 36.05 15.00
166 TYR CD2 35.98 30.82 36.64 15.00
166 TYR CE2 36.64 30.00 35.73 15.00
166 TYR CZ 37.96 30.23 35.43 15.00
166 TYR OH 38.59 29.45 34.50 15.00 166 TYR C 37.28 32.18 40.15 15.00
166 TYR 0 37.67 33.05 40.93 15.00
167 MET N 38.08 31.24 39.66 15.00
167 MET CA 39.44 31.09 40.16 15.00
167 MET CB 39.93 29.66 .39.91 15.00
167 MET CG 39.21 28.59 40.73 15.00
167 MET SD 37.76 27.96 39.92 15.00
167 MET CE 38.38 27.66 38.30 15.00
167 MET C 40.51 32.11 39.77 15.00
167 MET 0 41.15 32.69 40.66 15.00
168 THR N 40.72 32.34 38.48 15.00
168 THR CA 41.73 33.30 38.05 15.00
168 THR CB 41.79 33.47 36.51 15.00
168 THR OGl 40.50 33.82 36.01 15.00
168 THR CG2 42.21 32.18 35.84 15.00
168 THR C 41.59 34.64 38.79 15.00
168 THR 0 42.57 35.17 39.31 15.00
169 ASN N 40.36 35.14 38.92 15.00
169 ASN CA 40.14 36.39 39.64 15.00
169 ASN CB 38.66 36.76 •39.66 15.00
169 ASN CG 38.27 37.62 38.50 15.00
169 ASN ODl 37.59 37.17 37.58 15.00
169 ASN ND2 38.70 38.87 38.52 15.00
169 ASN C 40.64 36.31 41.07 15.00
169 ASN 0 40.97 37.33 41.65 15.00
170 ALA N 40.67 35.11 41.64 15.00
170 ALA CA 41.14 34.94 43.01 15.00
170 ALA CB 40.66 33.62 43.57 15.00
170 ALA C 42.67 35.02 43.12 15.00
170 ALA 0 43.22 35.37 44.17 15.00
171 PHE N 43.34 34.63 42.05 15.00
171 PHE CA 44.79 34.64 42.03 15.00
171 PHE CB 45.31 33.72 40.93 15.00
171 PHE CG 45.00 32.26 41.18 15.00
171 PHE CDl 45.01 31.75 •42.46 15.00
171 PHE CD2 44.71 31.41 40.12 15.00
171 PHE CEl 44.73 30.41 42.70 15.00 171 PHE CE2 44.44 30.06 40.35 15.00
171 PHE CZ 44.45 29.57 41.65 15.00
171 PHE C 45.31 36.06 41.88 15.00
171 PHE 0 46.16 36.49 42.68 15.00
172 GLN N 44.77 36.82 40.93 15.00
172 GLN CA 45.21 38.20 40.76 15.00
172 GLN CB 44.56 38.85 39.53 15.00
172 GLN CG 43.15 39.38 39.76 15.00
172 GLN CD 42.50 39.90 .38.48 15.00
172 GLN OEl 42.71 39.36 37.38 15.00
172 GLN NE2 41.69 40.95 38.61 15.00
172 GLN C 44.95 39.05 42.02 15.00
172 GLN 0 45.63 40.04 42.26 15.00
173 TYR N 43.97 38.65 42.83 15.00
173 TYR CA 43.67 39.38 44.05 15.00
173 TYR CB 42.40 38.87 44.74 15.00
173 TYR CG 42.31 39.27 46.21 15.00
173 TYR CDl 42.08 40.59 46.59 15.00
173 TYR CEl 42.08 40.96 47.94 15.00
173 TYR CD2 42.54 38.33 47.21 15.00
173 TYR CE2 42.55 38.69 48.56 15.00
173 TYR CZ 42.32 40.00 48.91 15.00
173 TYR OH 42.35 40.33 50.24 15.00
173 TYR C 44.83 39.26 .45.03 15.00
173 TYR 0 45.30 40.26 45.56 15.00
174 VAL N 45.29 38.04 45.27 15.00
174 VAL CA 46.39 37.82 46.20 15.00
174 VAL CB 46.76 36.32 46.27 15.00
174 VAL CGI 47.52 36.03 47.56 15.00
174 VAL CG2 45.51 35.45 46.17 15.00
174 VAL C 47.62 38.62 45.75 15.00
174 VAL 0 48.20 39.38 46.54 15.00
175 GLN N 47.98 38.48 44.48 15.00
175 GLN CA 49.12 39.18 43.88 15.00
175 GLN CB 49.09 38.97 42.36 15.00
175 GLN CG 49.97 39.89 41.54 15.00
175 GLN CD 49.72 39.72 40.06 15.00 175 GLN OEl 49.68 38.60 39.56 15.00
175 GLN NE2 49.51 40.82 39.36 15.00
175 GLN C 49.08 40.67 44.19 15.00
175 GLN 0 49.89 41.18 44.98 15.00
176 LYS N 48.09 41.36 43.63 15.00
176 LYS CA 47.95 42.79 43.83 15.00
176 LYS CB 46.79 43.32 42.99 15.00
176 LYS CG 47.01 43.12 41.48 15.00
176 LYS CD 45.77 43.45 40.65 15.00
176 LYS CE 45.42 44.92 40.74 15.00
176 LYS NZ 46.54 45.80 40.27 15.00
176 LYS C 47.76 43.14 45.30 15.00
176 LYS 0 48.07 44.26 45.72 15.00
177 ASN N 47.33 42.17 46.09 15.00
177 ASN CA 47.09 42.36 47.52 15.00
177 ASN CB 45.87 41.52 47.95 15.00
177 ASN CG 45.72 41.43 49.47 15.00
177 ASN ODl 45.26 42.37 50.13 15.00
177 ASN ND2 46.12 40.29 50.02 15.00
177 ASN C 48.30 42.05 48.41 15.00
177 ASN 0 48.28 42.38 49.60 15.00
178 ARG N 49.34 41.43 47.86 15.00
178 ARG CA 50.56 41.07 48.61 15.00
178 ARG CB 51.19 42.27 49.35 15.00
178 ARG CG 51.37 43.55 48.57 15.00
178 ARG CD 52.20 43.37 47.33 15.00
178 ARG NE 52.30 44.62 46.60 15.00
178 ARG CZ 52.31 44.71 45.28 15.00
178 ARG NHl 52.22 43.61 .44.53 15.00
178 ARG NH2 52.48 45.89 44.70 15.00
178 ARG C 50.34 39.99 49.65 15.00
178 ARG 0 50.95 40.04 50.71 15.00
179 GLY N 49.45 39.04 49.38 15.00
179 GLY CA 49.24 37.97 50.34 15.00
179 GLY C 47.84 37.46 50.53 15.00
179 GLY 0 46.89 37.97 49.95 15.00
180 ILE N 47.73 36.41 51.33 15.00 180 ILE CA 46.46 35.77 51.64 15.00
180 ILE CB 46.17 34.60 50.69 15.00
180 ILE CG2 47.30 33.58 50.74 15.00
180 ILE CGI 44.82 33.97 '51.03 15.00
180 ILE CDl 44.48 32.76 50.21 15.00
180 ILE C 46.57 35.27 53.06 15.00
180 ILE 0 47.49 34.54 53.41 15.00
181 ASP N 45.64 35.72 53.90 15.00
181 ASP CA 45.60 35.33 55.30 15.00
181 ASP CB 44.50 36.12 56.00 15.00
181 ASP CG 45.00 36.86 57.22 15.00
181 ASP ODl 44.78 38.09 57.30 15.00
181 ASP OD2 45.60 36.21 58.10 15.00
181 ASP C 45.32 33.84 55.39 15.00
181 ASP 0 44.86 33.23 54.43 15.00
182 SER N 45.60 33.24 56.53 15.00
182 SER CA 45.36 31.82 56.66 15.00
182 SER CB 46.51 31.16 57.42 15.00
182 SER OG 46.40 31.38 58.81 15.00
182 SER C 44.07 31.58 57.40 15.00
182 SER 0 43.68 32.37 58.25 15.00
183 GLU N 43.43 30.45 57.12 15.00
183 GLU CA 42.19 30.07 57.79 15.00
183 GLU CB 41.86 28.61 57.44 15.00
183 GLU CG 40.75 27.96 58.27 15.00
183 GLU CD 39.39 28.59 58.04 15.00
183 GLU OEl 39.09 29.02 56.90 15.00
183 GLU OE2 38.61 28.63 59.01 15.00
183 GLU C 42.42 30.21 59.28 15.00
183 GLU 0 41.61 30.74 60.02 15.00
184 ASP N 43.62 29.82 59.69 15.00
184 ASP CA 44.01 29.89 61.09 15.00
184 ASP CB 45.46 29.38 61.25 15.00
184 ASP CG 45.79 28.22 60.31 15.00
184 ASP ODl 45.54 27.05 60.71 15.00
184 ASP OD2 46.27 28.47 59.18 15.00
184 ASP C 43.90 31.32 61.60 15.00 184 ASP 0 43.51 31.54 62.74 15.00
185 ALA N 44.22 32.28 60.74 15.00
185 ALA CA 44.19 33.69 61.12 15.00
185 ALA CB 45.50 34.36 60.72 15.00
185 ALA C 43.02 34.48 60.56 15.00
185 ALA 0 42.92 35.69 60.78 15.00
186 TYR N 42.11 33.81 59.88 15.00
186 TYR CA 40.98 34.49 59.26 15.00
186 TYR CB 41.49 35.24 '58.02 15.00
186 TYR CG 40.53 36.17 57.35 15.00
186 TYR CDl 39.34 36.56 57.97 15.00
186 TYR CEl 38.48 37.45 57.34 15.00
186 TYR CD2 40.82 36.69 56.10 15.00
186 TYR CE2 39.96 37.58 55.47 15.00
186 TYR CZ 38.80 37.95 56.10 15.00
186 TYR OH 37.97 38.84 55.47 15.00
186 TYR C 40.02 33.39 58.86 15.00
186 TYR 0 40.00 32.96 57.71 15.00
187 PRO N 39.24 32.89 59.82 15.00
187 PRO CD 39.28 33.44 61.19 15.00
187 PRO CA 38.23 31.83 59.73 15.00
187 PRO CB 37.55 31.90 61.09 15.00
187 PRO CG 38.65 32.33 61.99 15.00
187 PRO C 37.20 31.93 58.62 15.00
187 PRO 0 36.96 32.99 58.07 15.00
188 TYR N 36.56 30.81 58.36 15.00
188 TYR CA 35.52 30.72 57.34 15.00
188 TYR CB 35.65 29.41 56.55 15.00
188 TYR CG 34.66 29.26 55.41 15.00
188 TYR CDl 34.74 30.08 54.27 15.00
188 TYR CEl 33.85 29.92 53.20 15.00
188 TYR CD2 33.66 28.28 55.45 15.00
188 TYR CE2 32.77 28.12 54.38 15.00
188 TYR CZ 32.88 28.94 53.26 15.00
188 TYR OH 32.03 28.77 52.21 15.00
188 TYR C 34.23 30.68 58.14 15.00
188 TYR 0 34.13 29.94 59.12 15.00 189 VAL N 33.26 31.49 57.76 15.00
189 VAL CA 32.00 31.48 58.50 15.00
189 VAL CB 31.63 32.87 59.03 15.00
189 VAL CGI 30.21 32.86 59.62 15.00
189 VAL CG2 32.65 33.28 60.11 15.00
189 VAL C 30.87 30.90 57.68 15.00
189 VAL 0 30.04 30.16 58.19 15.00
190 GLY N 30.85 31.21 56.39 15.00
190 GLY CA 29.81 30.71 55.52 15.00
190 GLY C 29.09 31.87 54.89 15.00
190 GLY 0 28.83 31.90 53.69 15.00
191 GLN N 28.81 32.88 55.70 15.00
191 GLN CA 28.11 34.05 •55.21 15.00
191 GLN CB 27.13 34.55 56.28 15.00
191 GLN CG 27.81 34.96 57.59 15.00
191 GLN CD 26.86 34.98 58.76 15.00
191 GLN OEl 26.34 33.93 59.18 15.00
191 GLN NE2 26.61 36.17 59.31 15.00
191 GLN C 29.12 35.13 54.90 15.00
191 GLN 0 29.97 35.43 55.73 15.00
192 GLU N 29.04 35.71 53.71 15.00
192 GLU CA 29.97 36.78 53.34 15.00
192 GLU CB 29.72 37.25 51.90 15.00
192 GLU CG 28.31 37.78 51.63 15.00
192 GLU CD 28.07 38.20 50.17 15.00
192 GLU OEl 27.23 39.09 49.94 15.00
192 GLU OE2 28.71 37.63 49.24 15.00
192 GLU C 29.84 37.94 •54.33 15.00
192 GLU 0 28.83 38.06 55.02 15.00
193 GLU N 30.90 38.73 54.45 15.00
193 GLU CA 30.90 39.90 55.33 15.00
193 GLU CB 31.14 39.51 56.79 15.00
193 GLU CG 32.18 38.42 57.04 15.00
193 GLU CD 32.27 38.04 58.52 15.00
193 GLU OEl 33.25 37.35 58.91 15.00
193 GLU OE2 31.38 38.45 59.31 15.00
193 GLU C 31.93 40.91 54.83 15.00 193 GLU 0 32.49 40.73 53.75 15.00
194 SER N 32.15 41.98 55.58 15.00
194 SER CA 33.12 42.98 55.15 15.00
194 SER CB 33.02 44.25 56.02 15.00
194 SER OG 31.89 45.03 55.66 15.00
194 SER C 34.56 42.47 55.13 15.00
194 SER 0 35.04 41.86 56.09 15.00
195 CYS N 35.24 42.70 54.00 15.00
195 CYS CA 36.63 42.28 53.84 15.00
195 CYS C 37.45 42.85 54.99 15.00
195 CYS 0 37.66 44.06 55.08 15.00
195 CYS CB 37.21 42.77 52.52 15.00
195 CYS SG 39.00 42.47 52.43 15.00
196 MET N 37.88 41.97 55.87 15.00
196 MET CA 38.66 42.37 57.02 15.00
196 MET CB 37.88 42.07 58.30 15.00
196 MET CG 37.23 40.70 58.31 15.00
196 MET SD 36.08 40.54 .59.66 15.00
196 MET CE 37.05 39.49 60.82 15.00
196 MET C 39.98 41.63 56.99 15.00
196 MET 0 40.27 40.79 57.85 15.00
197 TYR N 40.75 41.92 55.95 15.00
197 TYR CA 42.06 41.33 55.74 15.00
197 TYR CB 42.54 41.61 54.32 15.00
197 TYR CG 43.92 41.10 54.00 15.00
197 TYR CDl 44.75 41.78 53.12 15.00
197 TYR CEl 45.98 41.28 52.76 15.00
197 TYR CD2 44.38 39.89 54.52 15.00
197 TYR CE2 45.62 39.38 54.16 15.00
197 TYR CZ 46.40 40.08 53.28 15.00
197 TYR OH 47.57 39.52 52.82 15.00
197 TYR C 43.02 41.98 56.69 15.00
197 TYR 0 42.85 43.14 .57.03 15.00
198 ASN N 44.04 41.24 57.10 15.00
198 ASN CA 45.06 41.78 57.98 15.00
198 ASN CB 44.76 41.50 59.44 15.00
198 ASN CG 45.80 42.08 60.36 15.00 198 ASN ODl 46.82 42.59 59.89 15.00
198 ASN ND2 45.55 42.04 61.66 15.00
198 ASN C 46.41 41.20 57.63 15.00
198 ASN O 46.68 40.02 57.89 15.00
199 PRO N 47.31 42.02 57.10 15.00
199 PRO CD 47.07 43.46 56.88 15.00
199 PRO CA 48.66 41.66 56.70 15.00
199 PRO CB 49.28 43.01 '56.34 15.00
199 PRO CG 48.10 43.80 55.84 15.00
199 PRO C 49.49 40.91 57.76 15.00
199 PRO O 50.45 40.22 57.43 15.00
200 THR N 49.10 41.04 59.02 15.00
200 THR CA 49.82 40.36 60.09 15.00
200 THR CB 49.29 40.80 61.44 15.00
200 THR OGl 47.92 40.37 61.56 15.00
200 THR CG2 49.37 42.33 61.58 15.00
200 THR C 49.70 38.83 60.00 15.00
200 THR O 50.46 38.09 60.62 15.00
201 GLY N 48.68 38.38 59.27 15.00
201 GLY CA 48.47 36.96 59.10 15.00
201 GLY C 48.75 36.56 57.66 15.00
201 GLY O 48.15 35.63 57.15 15.00
202 LYS N 49.61 37.33 '56.99 15.00
202 LYS CA 49.98 37.05 55.61 15.00
202 LYS CB 51.10 38.02 55.17 15.00
202 LYS CG 51.29 38.17 53.66 15.00
202 LYS CD 52.46 39.10 53.34 15.00
202 LYS CE 52.21 40.54 53.79 15.00
202 LYS NZ 51.96 41.48 52.65 15.00
202 LYS C 50.52 35.64 55.69 15.00
202 LYS O 51.33 35.33 56.57 15.00
203 ALA N 50.08 34.77 54.79 15.00
203 ALA CA 50.52 33.38 54.86 15.00
203 ALA CB 49.35 32.49 55.25 15.00
203 ALA C 51.16 32.87 53.60 15.00
203 ALA O 52.05 32.03 53.65 15.00
204 ALA N 50.68 33.33 52.46 15.00 204 ALA CA 51.23 32.90 51.20 15.00
204 ALA CB 50.40 31.76 50.65 15.00
204 ALA C 51.17 34.09 50.27 15.00
204 ALA 0 50.71 35.16 50.66 15.00
205 LYS N 51.65 33.91 49.05 15.00
205 LYS CA 51.64 34.98 48.07 15.00
205 LYS CB 52.98 35.71 48.04 15.00
205 LYS CG 53.06 36.98 48.87 15.00
205 LYS CD 54.37 37.70 48.59 15.00
205 LYS CE 54.47 38.99 49.39 15.00
205 LYS NZ 55.68 39.79 49.04 15.00
205 LYS C 51.38 34.38 •46.71 15.00
205 LYS 0 51.31 33.18 46.55 15.00
206 CYS N 51.29 35.24 45.72 15.00
206 CYS CA 51.05 34.77 44.38 15.00
206 CYS CB 49.56 34.72 44.11 15.00
206 CYS SG 49.22 34.26 42.42 15.00
206 CYS C 51.68 35.81 43.53 15.00
206 CYS 0 51.64 36.98 43.86 15.00
207 ARG N 52.31 35.37 42.45 15.00
207 ARG CA 52.97 36.30 41.56 15.00
207 ARG CB 54.49 36.03 41.61 15.00
207 ARG CG 55.05 36.33 43.04 15.00
207 ARG CD 56.51 35.94 43.35 15.00
207 ARG NE 56.81 36.25 44.76 15.00
207 ARG CZ 57.91 36.88 45.20 15.00
207 ARG NHl 58.87 37.28 •44.36 15.00
207 ARG NH2 58.01 37.20 46.49 15.00
207 ARG C 52.37 36.22 40.16 15.00
207 ARG 0 52.85 36.86 39.22 15.00
208 GLY N 51.25 35.51 40.07 15.00
208 GLY CA 50.55 35.33 38.81 15.00
208 GLY C 49.86 33.97 38.78 15.00
208 GLY 0 49.80 33.27 39.79 15.00
209 TYR N 49.32 33.60 37.63 15.00
209 TYR CA 48.65 32.32 37.46 15.00
209 TYR CB 47.14 32.43 37.78 15.00 209 TYR CG 46.30 33.17 36.74 15.00
209 TYR CDl 45.95 32.54 •35.54 15.00
209 TYR CEl 45.27 33.23 34.53 15.00
209 TYR CD2 45.95 34.51 36.91 15.00
209 TYR CE2 45.27 35.20 35.90 15.00
209 TYR CZ 44.94 34.55 34.72 15.00
209 TYR OH 44.29 35.23 33.70 15.00
209 TYR C 48.85 31.91 36.01 15.00
209 TYR 0 49.27 32.72 35.18 15.00
210 ARG N 48.59 30.64 35.71 15.00
210 ARG CA 48.70 30.12 34.35 15.00
210 ARG CB 49.96 29.27 34.19 15.00
210 ARG CG 51.15 30.04 33.66 15.00
210 ARG CD 50.76 30.86 32.42 15.00
210 ARG NE 51.90 31.36 31.65 15.00
210 ARG CZ 53.02 31.88 32.16 15.00
210 ARG NHl 53.21 31.98 •33.47 15.00
210 ARG NH2 53.96 32.33 31.32 15.00
210 ARG C 47.48 29.26 34.05 15.00
210 ARG 0 47.02 28.47 34.88 15.00
211 GLU N 46.94 29.41 32.84 15.00
211 GLU CA 45.77 28.65 32.46 15.00
211 GLU CB 44.83 29.54 31.65 15.00
211 GLU CG 44.64 30.92 32.25 15.00
211 GLU CD 43.20 31.38 32.17 15.00
211 GLU OEl 42.37 30.81 32.91 15.00
211 GLU OE2 42.90 32.31 31.39 15.00
211 GLU C 46.17 27.46 31.63 15.00
211 GLU 0 46.84 27.61 30.61 15.00
212 ILE N 45.81 26.27 32.09 15.00
212 ILE CA 46.11 25.06 31.33 15.00
212 ILE CB 45.66 23.79 32.07 15.00
212 ILE CG2 45.75 22.59 31.16 15.00
212 ILE CGI 46.53 23.52 33.30 15.00
212 ILE CDl 46.30 24.44 34.45 15.00
212 ILE C 45.28 25.23 30.07 15.00
212 ILE 0 44.17 25.76 30.13 15.00 213 PRO N 45.82 24.86 28.90 15.00
213 PRO CD 47.16 24.29 28.66 15.00
213 PRO CA 45.09 25.00 27.64 15.00
213 PRO CB 45.89 24.12 26.68 15.00
213 PRO CG 47.28 24.36 27.14 15.00
213 PRO C 43.64 24.56 .27.74 15.00
213 PRO 0 43.30 23.67 28.53 15.00
214 GLU N 42.80 25.16 26.91 15.00
214 GLU CA 41.35 24.90 26.88 15.00
214 GLU CB 40.68 26.02 26.06 15.00
214 GLU CG 39.26 26.39 26.48 15.00
214 GLU CD 38.19 25.45 25.93 15.00
214 GLU OEl 37.01 25.61 26.34 15.00
214 GLU OE2 38.51 24.58 25.08 15.00
214 GLU C 40.94 23.52 26.33 15.00
214 GLU 0 41.21 23.20 25.18 15.00
215 GLY N 40.28 22.73 27.17 15.00
215 GLY CA 39.79 21.42 26.76 15.00
215 GLY C 40.76 20.25 26.77 15.00
215 GLY 0 40.41 19.13 27.15 15.00
216 ASN N 41.98 20.51 .26.32 15.00
216 ASN CA 43.03 19.50 26.24 15.00
216 ASN CB 44.27 20.10 25.58 15.00
216 ASN CG 45.47 19.19 25.64 15.00
216 ASN ODl 45.36 18.02 26.01 15.00
216 ASN ND2 46.63 19.73 25.30 15.00
216 ASN C 43.40 18.87 27.59 15.00
216 ASN 0 43.84 19.56 28.50 15.00
217 GLU N 43.29 17.55 27.67 15.00
217 GLU CA 43.59 16.83 28.91 15.00
217 GLU CB 42.71 15.60 29.05 15.00
217 GLU CG 41.25 15.91 29.21 15.00
217 GLU CD 40.42 14.67 29.07 15.00
217 GLU OEl 39.86 14.22 30.09 15.00
217 GLU OE2 40.34 14.14 27.93 15.00
217 GLU C 45.04 16.41 •29.12 15.00
217 GLU 0 45.47 16.27 30.26 15.00 218 LYS N 45.77 16.14 28.04 15.00
218 LYS CA 47.17 15.75 28.17 15.00
218 LYS CB 47.80 15.49 26.80 15.00
218 LYS CG 47.16 14.35 26.04 15.00
218 LYS CD 47.73 14.24 24.62 15.00
218 LYS CE 49.20 13.75 24.61 15.00
218 LYS NZ 49.81 13.60 23.23 15.00
218 LYS C 47.89 16.92 28.85 15.00
218 LYS 0 48.79 16.73 29.67 15.00
219 ALA N 47.45 18.13 28.49 15.00
219 ALA CA 48.01 19.34 .29.04 15.00
219 ALA CB 47.35 20.53 28.40 15.00
219 ALA C 47.73 19.33 30.53 15.00
219 ALA 0 48.61 19.63 31.34 15.00
220 LEU N 46.51 18.94 30.89 15.00
220 LEU CA 46.14 18.91 32.29 15.00
220 LEU CB 44.65 18.56 32.44 15.00
220 LEU CG 44.05 18.37 33.84 15.00
220 LEU CDl 44.26 19.58 34.76 15.00
220 LEU CD2 42.58 18.06 33.71 15.00
220 LEU C 47.02 17.91 33.02 15.00
220 LEU 0 47.55 18.22 34.08 15.00
221 LYS N 47.24 16.76 32.40 15.00
221 LYS CA 48.06 15.67 32.97 15.00
221 LYS CB 48.19 14.53 31.95 15.00
221 LYS CG 49.20 13.47 .32.33 15.00
221 LYS CD 49.35 12.44 31.22 15.00
221 LYS CE 50.40 11.43 31.59 15.00
221 LYS NZ 50.20 10.97 33.00 15.00
221 LYS C 49.45 16.11 33.37 15.00
221 LYS 0 49.92 15.76 34.45 15.00
222 ARG N 50.12 16.82 32.48 15.00
222 ARG CA 51.47 17.32 32.72 15.00
222 ARG CB 52.06 17.83 31.42 15.00
222 ARG CG 52.01 16.80 30.32 15.00
222 ARG CD 53.17 15.86 30.42 15.00
222 ARG NE 54.37 16.47 29.86 15.00 222 ARG CZ 54.74 16.35 28.59 15.00
222 ARG NHl 54.00 15.63 27.74 15.00
222 ARG NH2 55.86 16.92 28.16 15.00
222 ARG C 51.41 18.43 33.76 15.00
222 ARG 0 52.15 18.39 34.75 15.00
223 ALA N 50.48 19.36 33.58 15.00
223 ALA CA 50.29 20.50 34.48 15.00
223 ALA CB 48.96 21.16 34.19 15.00
223 ALA C 50.33 20.07 35.94 15.00
223 ALA 0 51.06 20.63 36.74 15.00
224 VAL N 49.56 19.04 36.24 15.00
224 VAL CA 49.47 18.50 37.58 15.00
224 VAL CB 48.45 17.31 37.65 15.00
224 VAL CGI 48.43 16.70 39.04 15.00
224 VAL CG2 47.05 17.77 37.26 15.00
224 VAL C 50.84 18.03 38.04 15.00
224 VAL 0 51.42 18.62 38.94 15.00
225 ALA N 51.37 17.00 37.38 15.00
225 ALA CA 52.66 16.43 37.76 15.00
225 ALA CB 52.92 15.17 36.97 15.00
225 ALA C 53.87 17.36 37.66 15.00
225 ALA 0 54.88 17.13 38.32 15.00
226 ARG N 53.78 18.41 36.85 15.00
226 ARG CA 54.90 19.33 36.70 15.00
226 ARG CB 54.95 19.86 35.26 15.00
226 ARG CG 56.31 20.45 34.85 15.00
226 ARG CD 56.38 20.87 33.36 15.00
226 ARG NE 56.18 19.76 32.40 15.00
226 ARG CZ 56.94 18.66 32.34 15.00
226 ARG NHl 57.95 18.49 33.18 15.00
226 ARG NH2 56.67 17.72 31.43 15.00
226 ARG C 54.84 20.49 37.69 15.00
226 ARG 0 55.89 20.95 38.15 15.00
227 VAL N 53.63 20.93 38.03 15.00
227 VAL CA 53.45 22.06 38.94 15.00
227 VAL CB 52.54 23.12 38.31 15.00
227 VAL CGI 52.50 24.36 39.19 15.00 227 VAL CG2 53.01 23.45 36.92 15.00
227 VAL C 52.87 21.76 40.33 15.00
227 VAL O 53.17 22.46 41.29 15.00
228 GLY N 51.99 20.78 40.41 15.00
228 GLY CA 51.37 20.44 41.68 15.00
228 GLY C 49.87 20.36 .41.46 15.00
228 GLY O 49.45 20.39 40.30 15.00
229 PRO N 49.03 20.27 42.51 15.00
229 PRO CD 49.35 20.33 43.95 15.00
229 PRO CA 47.58 20.19 42.31 15.00
229 PRO CB 47.04 20.14 43.74 15.00
229 PRO CG 48.06 20.86 44.52 15.00
229 PRO C 47.05 21.39 41.55 15.00
229 PRO O 47.31 22.54 41.92 15.00
230 VAL N 46.34 21.13 40.46 15.00
230 VAL CA 45.75 22.19 39.65 15.00
230 VAL CB 45.95 21.89 38.16 15.00
230 VAL CGI 45.22 22.89 37.30 15.00
230 VAL CG2 47.40 21.90 37.84 15.00
230 VAL C 44.25 22.37 39.94 15.00
230 VAL O 43.59 21.42 ■40.37 15.00
231 SER N 43.74 23.58 39.76 15.00
231 SER CA 42.33 23.89 39.96 15.00
231 SER CB 42.17 25.38 40.29 15.00
231 SER OG 42.60 25.66 41.61 15.00
231 SER C 41.54 23.56 38.69 15.00
231 SER O 42.02 23.82 37.58 15.00
232 VAL N 40.35 22.96 38.84 15.00
232 VAL CA 39.50 22.62 37.69 15.00
232 VAL CB 39.65 21.13 37.25 15.00
232 VAL CGI 41.00 20.91 36.59 15.00
232 VAL CG2 39.47 20.20 38.43 15.00
232 VAL C 38.01 22.93 37.92 15.00
232 VAL O 37.59 23.17 39.06 15.00
233 ALA N 37.24 22.97 36.84 15.00
233 ALA CA 35.81 23.26 36.88 15.00
233 ALA CB 35.51 24.52 36.09 15.00 233 ALA C 35,.12 22,.07 36,.25 15,.00
233 ALA 0 35. .63 21. ,51 35, .29 15, .00
234 ILE N 33, .95 21. .70 36, .77 15. .00
234 ILE CA 33. .20 20, .54 36, .28 15, .00
234 ILE CB 33. .48 19, .29 37, .16 15. .00
234 ILE CG2 34, .83 18. .69 36. .84 15. .00
234 ILE CGI 33. .34 19, .68 38. .65 15. .00
234 ILE CDl 33. .22 18, .53 39, .59 15. .00
234 ILE C 31, .69 20. .74 36, .29 15, .00
234 ILE 0 31. .19 21. .79 36, .70 15. .00
235 ASP N 30. .97 19, .68 35, .90 15. .00
235 ASP CA 29, .52 19. .64 35, .85 15, .00
235 ASP CB 29, .06 19. .03 34, .53 15. .00
235 ASP CG 27. .57 19, .14 34, .32 15, .00
235 ASP ODl 26, .80 19. .01 35, .29 15. .00
235 ASP OD2 27. .17 19. .35 33, .15 15, .00
235 ASP C 29, .05 18, .74 36, .99 15. .00
235 ASP 0 28, .88 17. .54 36, .80 15. .00
236 ALA N 28. .83 19. .32 38, .16 15, .00
236 ALA CA 28, .40 18. .55 39, .32 15. .00
236 ALA CB 29. .13 19. .06 40, .56 15. .00
236 ALA C 26. .88 18, .61 39. .53 15. .00
236 ALA 0 26, .39 18. .67 40. .67 15, .00
237 SER N 26. .12 18. .60 38. .43 15. .00
237 SER CA 24. .66 18. .64 38, .52 15, .00
237 SER CB 24. .05 19. .01 37, .17 15, .00
237 SER OG 24. .41 20. .33 36, .81 15. ,00
237 SER C 24. .16 17. .27 38, .93 15, ,00
237 SER 0 23. .69 17. .09 40, .05 15. ,00
238 LEU N 24, .36 16. .31 38. .05 15. ,00
238 LEU CA 23. .94 14, .92 38, .23 15. .00
238 LEU CB 24. .85 14. .01 37. .41 15. ,00
238 LEU CG 24, .71 14, .03 35, .89 15, .00
238 LEU CDl 24. .06 15. .31 35. .38 15. ,00
238 LEU CD2 26, .08 13, .85 35. .28 15. ,00
238 LEU C 23, .82 14. .36 39. .65 15. ,00
238 LEU 0 24, .67 14, .62 40. .51 15. ,00 239 THR N 22.78 13.58 39.89 15.00
239 THR CA 22.57 12.97 41.20 15.00
239 THR CB 21.14 12.40 41.34 15.00
239 THR OGl 20.58 12.17 40.04 15.00
239 THR CG2 20.25 13.37 42.13 15.00
239 THR C 23.59 11.85 41.41 15.00
239 THR 0 24.04 11.62 42.54 15.00
240 SER N 23.97 11.18 40.33 15.00
240 SER CA 24.95 10.12 40.39 15.00
240 SER CB 25.24 9.60 38.99 15.00
240 SER OG 25.34 10.66 38.08 15.00
240 SER C 26.22 10.69 41.02 15.00
240 SER 0 26.97 9.99 41.69 15.00
241 PHE N 26.45 11.98 40.79 15.00
241 PHE CA 27.58 12.67 •41.37 15.00
241 PHE CB 27.83 13.99 40.63 15.00
241 PHE CG 29.04 14.74 41.12 15.00
241 PHE CDl 30.17 14.85 40.32 15.00
241 PHE CD2 29.03 15.38 42.35 15.00
241 PHE CEl 31.27 15.59 40.74 15.00
241 PHE CE2 30.12 16.12 42.77 15.00
241 PHE CZ 31.24 16.22 41.96 15.00
241 PHE C 27.26 12.95 42.84 15.00
241 PHE 0 27.92 12.42 43.73 15.00
242 GLN N 26.20 13.71 43.09 15.00
242 GLN CA 25.78 14.08 44.44 15.00
242 GLN CB 24.36 14.62 44.42 15.00
242 GLN CG 23.88 15.20 45.73 15.00
242 GLN CD 22.37 15.17 45.88 15.00
242 GLN OEl 21.83 15.59 46.89 15.00
242 GLN NE2 21.68 14.65 44.87 15.00
242 GLN C 25.86 12.98 45.49 15.00
242 GLN 0 26.05 13.27 46.69 15.00
243 PHE N 25.70 11.73 45.06 15.00
243 PHE CA 25.72 10.58 45.97 15.00
243 PHE CB 24.45 9.74 45.79 15.00
243 PHE CG 23.22 10.44 46.23 15.00 243 PHE CDl 22.95 10.59 47.58 15.00
243 PHE CD2 22.36 11.00 45.30 15.00
243 PHE CEl 21.85 11.31 48.01 15.00
243 PHE CE2 21.26 11.72 45.72 15.00
243 PHE CZ 21.00 11.88 47.08 15.00
243 PHE C 26.93 9.66 45.86 15.00
243 PHE 0 26.97 8.63 46.54 15.00
244 TYR N 27.88 10.00 44.99 15.00
244 TYR CA 29.09 9.20 44.81 15.00
244 TYR CB 30.17 10.02 44.09 15.00
244 TYR CG 31.52 9.34 44.06 15.00
244 TYR CDl 31.86 8.43 43.05 15.00
244 TYR CEl 33.07 7.72 43.09 15.00
244 TYR CD2 32.42 9.52 45.10 15.00
244 TYR CE2 33.62 8.82 45.16 15.00
244 TYR CZ 33.94 7.92 44.16 15.00
244 TYR OH 35.12 7.20 44.29 15.00
244 TYR C 29.63 8.64 46.13 15.00
244 TYR 0 29.53 9.28 47.18 15.00
245 SER N 30.20 7.44 46.06 15.00
245 SER CA 30.78 6.78 47.22 15.00
245 SER CB 29.88 5.63 47.69 15.00
245 SER OG 30.30 5.06 48.92 15.00
245 SER C 32.19 6.25 46.92 15.00
245 SER 0 33.16 6.62 47.60 15.00
246 LYS N 32.31 5.39 45.91 15.00
246 LYS CA 33.61 4.81 45.57 15.00
246 LYS CB 33.82 3.50 46.33 15.00
246 LYS CG 34.29 3.71 47.75 15.00
246 LYS CD 33.63 2.74 48.71 15.00
246 LYS CE 34.06 3.03 50.15 15.00
246 LYS NZ 33.94 4.48 50.54 15.00
246 LYS C 33.71 4.56 44.08 15.00
246 LYS 0 32.73 4.75 .43.35 15.00
247 GLY N 34.90 4.18 43.62 15.00
247 GLY CA 35.09 3.90 42.21 15.00
247 GLY C 35.36 5.08 41.29 15.00 247 GLY 0 35.42 6.22 41.72 15.00
248 VAL N 35.53 4.77 40.02 15.00
248 VAL CA 35.81 5.78 39.01 15.00
248 VAL CB 36.66 5.18 37.87 15.00
248 VAL CGI 36.96 6.23 36.82 15.00
248 VAL CG2 37.96 4.62 38.43 15.00
248 VAL C 34.47 6.26 38.47 15.00
248 VAL 0 33.75 5.51 37.82 15.00
249 TYR N 34.15 7.51 •38.77 15.00
249 TYR CA 32.90 8.12 38.37 15.00
249 TYR CB 32.58 9.29 39.30 15.00
249 TYR CG 31.31 10.02 38.94 15.00
249 TYR CDl 30.12 9.78 39.63 15.00
249 TYR CEl 28.94 10.43 39.27 15.00
249 TYR CD2 31.28 10.95 37.90 15.00
249 TYR CE2 30.11 11.59 37.54 15.00
249 TYR CZ 28.95 11.34 38.22 15.00
249 TYR OH 27.80 12.00 37.86 15.00
249 TYR C 32.80 8.59 36.92 15.00
249 TYR 0 33.42 9.56 36.52 15.00
250 TYR N 31.91 7.95 36.18 15.00
250 TYR CA 31.66 8.32 34.80 15.00
250 TYR CB 32.39 7.39 33.84 15.00
250 TYR CG 32.13 7.70 ■32.38 15.00
250 TYR CDl 32.16 9.01 31.91 15.00
250 TYR CEl 31.94 9.28 30.57 15.00
250 TYR CD2 31.89 6.67 31.48 15.00
250 TYR CE2 31.68 6.93 30.14 15.00
250 TYR CZ 31.70 8.23 29.69 15.00
250 TYR OH 31.47 8.48 28.36 15.00
250 TYR C 30.17 8.21 34.56 15.00
250 TYR 0 29.58 7.14 34.75 15.00
251 ASP N 29.54 9.33 34.23 15.00
251 ASP CA 28.12 9.34 33.96 15.00
251 ASP CB 27.37 10.20 34.98 15.00
251 ASP CG 25.84 10.11 34.84 15.00
251 ASP ODl 25.34 9.43 33.91 15.00 251 ASP OD2 25.13 10.73 35.65 15.00
251 ASP C 27.98 9.93 32.57 15.00
251 ASP 0 28.34 11.08 32.35 15.00
252 GLU N 27.39 9.15 31.67 15.00
252 GLU CA 27.18 9.55 30.28 15.00
252 GLU CB 26.55 8.41 29.45 15.00
252 GLU CG 26.94 6.97 29.83 15.00
252 GLU CD 26.21 6.45 31.07 15.00
252 GLU OEl 25.02 6.81 31.27 15.00
252 GLU OE2 26.82 5.71 31.87 15.00
252 GLU C 26.27 10.78 30.18 15.00
252 GLU 0 25.85 11.15 29.08 15.00
253 SER N 25.95 11.39 31.32 15.00
253 SER CA 25.06 12.53 '31.35 15.00
253 SER CB 23.92 12.21 32.30 15.00
253 SER OG 23.50 10.85 32.13 15.00
253 SER C 25.71 13.86 31.72 15.00
253 SER 0 25.08 14.92 31.61 15.00
254 CYS N 26.95 13.81 32.21 15.00
254 CYS CA 27.69 15.02 32.58 15.00
254 CYS C 27.77 15.85 31.30 15.00
254 CYS 0 27.93 15.30 30.21 15.00
254 CYS CB 29.12 14.65 33.04 15.00
254 CYS SG 29.66 15.23 34.70 15.00
255 ASN N 27.60 17.16 31.41 15.00
255 ASN CA 27.69 18.00 30.23 15.00
255 ASN CB 26.41 18.79 30.01 15.00
255 ASN CG 26.54 19.76 28.85 15.00
255 ASN ODl 26.34 20.96 29.02 15.00
255 ASN ND2 26.94 19.25 27.69 15.00
255 ASN C 28.86 18.98 30.26 15.00
255 ASN 0 28.94 19.82 31.14 15.00
256 SER N 29.71 18.89 29.24 15.00
256 SER CA 30.88 19.75 29.10 15.00
256 SER CB 31.66 19.38 27.83 15.00
256 SER OG 32.00 18.00 27.83 15.00
256 SER C 30.57 21.25 29.07 15.00 256 SER 0 31,,42 22,.06 29,.45 15..00
257 ASP N 29, .39 21, .63 28. .59 15. .00
257 ASP CA 29. .00 23, .04 28, .53 15. .00
257 ASP CB 27. .96 23. .26 27. .44 15. .00
257 ASP CG 28. .50 23. .00 26. .05 15. .00
257 ASP ODl 28. .50 21, .82 25. .63 15. .00
257 ASP OD2 28. .91 23. .96 25. .36 15. .00
257 ASP C 28, .47 23, .61 29, .85 15, .00
257 ASP 0 28. .51 24, .82 30, .07 15. .00
258 ASN N 27. .95 22. .76 30, .74 15. .00
258 ASN CA 27. .41 23, .23 32, .01 15. .00
258 ASN CB 26, .15 22. .43 32, .39 15. .00
258 ASN CG 25, .57 22, .82 33, .75 15, .00
258 ASN ODl 25, .50 24, .00 34, .10 15. .00
258 ASN ND2 25, .15 21, .83 34. .53 15. .00
258 ASN C 28, .45 23, .14 33, .14 15. .00
258 ASN 0 28, .27 22, .37 34, .09 15, .00
259 LEU N 29, .53 23, .90 33, .03 15, .00
259 LEU CA 30, .56 23, .89 •34, .06 15. ,00
259 LEU CB 31, .79 24, .65 33. .58 15, .00
259 LEU CG 32, .55 24, .17 32, .34 15, .00
259 LEU CDl 33, .69 25, .13 32, .06 15, ,00
259 LEU CD2 33 .08 22 .77 32, .57 15, .00
259 LEU C 29, .99 24, .59 35, .28 15, .00
259 LEU 0 29 .76 25 .80 35, .25 15. .00
260 ASN N 29, .75 23, .87 36, .36 15. ,00
260 ASN CA 29, .17 24, .51 37, .52 15. .00
260 ASN CB 27 .70 24, .15 37, .61 15. .00
260 ASN CG 27, .47 22, .66 37, .68 15. ,00
260 ASN ODl 28 .02 21 .99 38, .54 15. .00
260 ASN ND2 26, .68 22, .14 36, .75 15, ,00
260 ASN C 29, .82 24, .27 38, .87 15. .00
260 ASN 0 29, .30 24 .73 39, .88 15, ,00
261 HIS N 30, .91 23, .52 •38. .92 15, ,00
261 HIS CA 31 .57 23 .30 40. .20 15, .00
261 HIS CB 31, .25 21, .91 40. .75 15. .00
261 HIS CG 31 .51 21 .75 42, .23 15. ,00 261 HIS CD2 31.94 20.69 42.95 15.00
261 HIS NDl 31.30 22.77 43.13 15.00
261 HIS CEl 31.59 22.34 44.35 15.00
261 HIS NE2 31.98 21.08 44.26 15.00
261 HIS C 33.07 23.51 40.04 15.00
261 HIS 0 33.61 23.35 38.95 15.00
262 ALA N 33.72 23.92 41.14 15.00
262 ALA CA 35.15 24.16 41.18 15.00
262 ALA CB 35.41 25.58 41.62 15.00
262 ALA C 35.81 23.18 42.16 15.00
262 ALA 0 35.49 23.18 43.35 15.00
263 VAL N 36.75 22.39 41.66 15.00
263 VAL CA 37.44 21.39 42.46 15.00
263 VAL CB 36.85 19.99 42.18 15.00
263 VAL CGI 35.47 19.89 42.73 15.00
263 VAL CG2 36.82 19.71 40.69 15.00
263 VAL C 38.97 21.34 42.25 15.00
263 VAL 0 39.51 21.84 41.26 15.00
264 LEU N 39.66 20.70 43.18 15.00
264 LEU CA 41.11 20.58 43.16 15.00
264 LEU CB 41.65 20.69 44.59 15.00
264 LEU CG 43.12 21.07 44.77 15.00
264 LEU CDl 43.26 22.53 44.44 15.00
264 LEU CD2 43.59 20.81 46.19 15.00
264 LEU C 41.59 19.25 42.56 15.00
264 LEU 0 41.33 18.20 43.14 15.00
265 ALA N 42.28 19.30 41.43 15.00
265 ALA CA 42.81 18.09 40.83 15.00
265 ALA CB 43.08 18.31 39.37 15.00
265 ALA C 44.12 17.84 41.56 15.00
265 ALA 0 45.02 18.66 41.48 15.00
266 VAL N 44.22 16.74 42.30 15.00
266 VAL CA 45.45 16.43 43.03 15.00
266 VAL CB 45.21 16.32 44.55 15.00
266 VAL CGI 44.51 17.56 45.07 15.00
266 VAL CG2 44.42 15.07 44.88 15.00
266 VAL C 46.21 15.19 42.57 15.00 266 VAL O 47.11 14.72 43.27 15.00
267 GLY N 45.85 14.65 41.42 15.00 267 GLY CA 46.54 13.48 40.92 15.00 267 GLY C 45.81 12.84 39.77 15.00
267 GLY O 44.81 13.37 39.29 15.00
268 TYR N 46.32 11.71 39.29 15.00 268 TYR CA 45.70 10.99 38.19 15.00 268 TYR CB 46.02 11.65 36.85 15.00 268 TYR CG 47.49 11.81 36.57 15.00 268 TYR CDl 48.25 10.74 36.09 15.00 268 TYR CEl 49.58 10.89 35.79 15.00 268 TYR CD2 48.13 13.03 36.74 15.00 268 TYR CE2 49.48 13.19 .36.44 15.00 268 TYR CZ 50.19 12.11 35.96 15.00 268 TYR OH 51.50 12.25 35.62 15.00 268 TYR C 46.17 9.55 38.24 15.00
268 TYR 0 46.87 9.17 39.19 15.00
269 GLY N 45.75 8.72 37.29 15.00 269 GLY CA 46.19 7.34 37.30 15.00 269 GLY C 45.08 6.30 37.24 15.00
269 GLY 0 43.92 6.58 37.55 15.00
270 ILE N 45.47 5.08 36.88 15.00 270 ILE CA 44.56 3.96 36.74 15.00 270 ILE CB 45.22 2.86 35.89 15.00 270 ILE CG2 44.36 1.61 35.83 15.00 270 ILE CGI 45.43 3.39 34.48 15.00 270 ILE CDl 45.60 2.31 33.44 15.00 270 ILE C 44.09 3.43 -38.10 15.00
270 ILE 0 44.69 3.75 39.13 15.00
271 GLN N 42.96 2.71 38.10 15.00 271 GLN CA 42.38 2.14 39.33 15.00 271 GLN CB 41.10 2.86 39.75 15.00 271 GLN CG 40.45 2.30 41.01 15.00 271 GLN CD 41.33 2.44 42.25 15.00 271 GLN OEl 40.96 3.12 43.21 15.00 271 GLN NE2 42.49 1.80 42.23 15.00 271 GLN C 42.07 0.65 39.23 15.00 271 GLN 0 42.72 -0.16 39.88 15.00
272 LYS N 41.03 0.31 38.47 15.00
272 LYS CA 40.67 -1.10 .38.30 15.00
272 LYS CB 39.32 -1.37 38.96 15.00
272 LYS CG 39.13 -2.79 39.44 15.00
272 LYS CD 39.79 -3.05 40.79 15.00
272 LYS CE 39.57 -4.51 41.20 15.00
272 LYS NZ 40.01 -4.82 42.59 15.00
272 LYS C 40.60 -1.36 36.80 15.00
272 LYS 0 39.81 -2.17 36.33 15.00
273 GLY N 41.48 -0.70 36.06 15.00
273 GLY CA 41.50 -0.86 34.62 15.00
273 GLY C 41.01 0.42 33.97 15.00
273 GLY 0 41.00 0.56 32.74 15.00
274 ASN N 40.62 1.37 34.81 15.00
274 ASN CA 40.11 2.65 34.34 15.00
274 ASN CB 38.74 2.96 34.97 15.00
274 ASN CG 37.76 1.82 ■34.81 15.00
274 ASN ODl 37.24 1.58 33.73 15.00
274 ASN ND2 37.49 1.11 35.90 15.00
274 ASN C 41.08 3.73 34.80 15.00
274 ASN 0 41.52 3.71 35.96 15.00
275 LYS N 41.43 4.63 33.89 15.00
275 LYS CA 42.32 5.74 34.22 15.00
275 LYS CB 42.98 6.29 32.96 15.00
275 LYS CG 43.74 5.26 32.16 15.00
275 LYS CD 44.26 5.91 30.91 15.00
275 LYS CE 44.42 4.88 29.82 15.00
275 LYS NZ 44.59 5.58 28.51 15.00
275 LYS C 41.43 6.80 34.83 15.00
275 LYS 0 40.43 7.19 34.23 15.00
276 HIS N 41.82 7.31 35.99 15.00
276 HIS CA 41.01 8.30 •36.70 15.00
276 HIS CB 40.50 7.68 38.01 15.00
276 HIS CG 41.58 7.33 39.00 15.00
276 HIS CD2 42.30 8.10 39.86 15.00
276 HIS NDl 41.95 6.03 39.27 15.00 276 HIS CEl 42,.82 6..00 40..26 15..00
276 HIS NE2 43. .05 7. .25 40. .63 15. .00
276 HIS C 41, .73 9, .59 37. .01 15. .00
276 HIS 0 42. .92 9. .73 36, .73 15, .00
277 TRP N 40, .98 10. .53 37, .57 15. .00
277 TRP CA 41, ,52 11. .81 37. .99 15. .00
277 TRP CB 40. .75 12, .96 37. .34 15, .00
277 TRP CG 41, .22 13, .34 35, .99 15. .00
277 TRP CD2 42, .48 13, .95 35, .66 15. .00
277 TRP CE2 42, .48 14, .17 34. .27 15. .00
277 TRP CE3 43, .60 14, .33 36, .41 15. .00
277 TRP CDl 40. .54 13, .22 34, .81 15. .00
277 TRP NEl 41. .29 13, .71 33. .77 15. .00
277 TRP CZ2 43. .57 14, .74 33. .61 15. .00
277 TRP CZ3 44, .67 14 .90 35. .75 15. .00
277 TRP CH2 44, .65 15, .11 34, .36 15. .00
277 TRP C 41. .26 11, .82 39. .49 15, .00
277 TRP 0 40, .31 11 .17 39, .94 15. .00
278 ILE N 42, .12 12, .46 40, .29 15. .00
278 ILE CA 41. .86 12, .52 41. .73 15, .00
278 ILE CB 43. .07 12, .15 42. .58 15. .00
278 ILE CG2 42, .63 11 .96 44, .03 15, .00
278 ILE CGI 43, .70 10, .87 42, .05 15, .00
278 ILE CDl 44. .89 10, .39 '42. .85 15. .00
278 ILE C 41. ,44 13, .94 42. ,05 15. ,00
278 ILE 0 42, .23 14 .87 41, .94 15, .00
279 ILE N 40, .17 14, .10 42. .41 15. .00
279 ILE CA 39. .63 15, .42 42. .69 15. ,00
279 ILE CB 38. .48 15, .74 41, .71 15. ,00
279 ILE CG2 37, .57 16, .79 42. .28 15. .00
279 ILE CGI 39. .07 16, .16 40. .35 15. ,00
279 ILE CDl 38. .05 16, .69 39. .36 15. .00
279 ILE C 39, .17 15, .62 44. .13 15. ,00
279 ILE 0 38. .55 14, .74 44, .73 15. .00
280 LYS N 39. .51 16, .79 44, .66 15. ,00
280 LYS CA 39, ,17 17. .17 46. ,01 15. ,00
280 LYS CB 40. .34 17, .93 46. .63 15. ,00 280 LYS CG 40.12 18.37 48.07 15.00
280 LYS CD 41.34 19.11 48.58 15.00
280 LYS CE 41.13 19.63 49.98 15.00
280 LYS NZ 42.34 20.34 50.48 15.00
280 LYS C 37.97 18.09 45.95 15.00
280 LYS 0 37.95 19.07 45.20 15.00
281 ASN N 36.94 17.76 46.74 15.00
281 ASN CA 35.74 18.57 46.76 15.00
281 ASN CB 34.48 17.70 46.73 15.00
281 ASN CG 33.38 18.32 45.91 15.00
281 ASN ODl 32.82 19.36 46.27 15.00
281 ASN ND2 33.09 17.72 44.77 15.00
281 ASN C 35.73 19.48 •47.98 15.00
281 ASN 0 36.70 19.50 48.74 15.00
282 SER N 34.62 20.20 48.15 15.00
282 SER CA 34.44 21.13 49.26 15.00
282 SER CB 34.30 22.55 48.70 15.00
282 SER OG 34.88 22.65 47.41 15.00
282 SER C 33.19 20.80 50.05 15.00
282 SER 0 32.67 21.65 50.76 15.00
283 TRP N 32.71 19.57 49.93 15.00
283 TRP CA 31.49 19.17 50.65 15.00
283 TRP CB 30.50 18.46 49.71 15.00
283 TRP CG 30.00 19.32 48.58 15.00
283 TRP CD2 29.39 18.88 47.37 15.00
283 TRP CE2 29.11 20.02 46.61 15.00
283 TRP CE3 29.06 17.62 46.87 15.00
283 TRP CDl 30.06 20.69 48.50 15.00
283 TRP NEl 29.52 21.11 47.32 15.00
283 TRP CZ2 28.51 19.94 45.35 15.00
283 TRP CZ3 28.47 17.54 45.62 15.00
283 TRP CH2 28.19 18.70 44.88 15.00
283 TRP C 31.75 18.34 51.92 15.00
283 TRP 0 31.01 17.41 52.22 15.00
284 GLY N 32.76 18.73 52.68 15.00
284 GLY CA 33.08 18.03 53.91 15.00
284 GLY C 33.53 16.59 53.76 15.00 284 GLY 0 33.58 16.05 52.66 15.00
285 GLU N 33.88 15.99 ■54.89 15.00
285 GLU CA 34.34 14.61 54.97 15.00
285 GLU CB 34.76 14.28 56.41 15.00
285 GLU CG 33.79 14.81 57.52 15.00
285 GLU CD 33.04 13.71 58.29 15.00
285 GLU OEl 33.21 12.51 57.98 15.00
285 GLU OE2 32.28 14.05 59.23 15.00
285 GLU C 33.33 13.56 54.51 15.00
285 GLU 0 33.65 12.70 53.68 15.00
286 ASN N 32.13 13.60 55.09 15.00
286 ASN CA 31.05 12.66 54.80 15.00
286 ASN CB 29.89 12.86 55.78 15.00
286 ASN CG 30.14 12.22 57.12 15.00
286 ASN ODl 29.89 12.83 58.16 15.00
286 ASN ND2 30.63 10.98 57.12 15.00
286 ASN C 30.47 12.72 •53.38 15.00
286 ASN 0 29.25 12.86 53.23 15.00
287 TRP N 31.31 12.62 52.36 15.00
287 TRP CA 30.83 12.63 50.98 15.00
287 TRP CB 30.76 14.06 50.45 15.00
287 TRP CG 30.33 14.12 49.00 15.00
287 TRP CD2 31.18 14.10 47.84 15.00
287 TRP CE2 30.35 14.15 46.71 15.00
287 TRP CE3 32.57 14.04 47.66 15.00
287 TRP CDl 29.05 14.18 48.53 15.00
287 TRP NEl 29.05 14.21 47.16 15.00
287 TRP CZ2 30.85 14.14 45.40 15.00
287 TRP CZ3 33.07 14.03 46.35 15.00
287 TRP CH2 32.21 14.08 45.24 15.00
287 TRP C 31.81 11.83 50.15 15.00
287 TRP 0 33.01 11.85 ■50.44 15.00
288 GLY N 31.33 11.14 49.12 15.00
288 GLY CA 32.22 10.36 48.28 15.00
288 GLY C 33.18 9.53 49.11 15.00
288 GLY 0 32.85 9.17 50.24 15.00
289 ASN N 34.39 9.29 48.60 15.00 289 ASN CA 35.36 8.48 49.34 15.00
289 ASN CB 36.28 7.74 48.38 15.00
289 ASN CG 37.06 6.65 49.07 15.00
289 ASN ODl 37.13 6.61 50.30 15.00
289 ASN ND2 37.63 5.75 48.28 15.00
289 ASN C 36.20 9.30 50.31 15.00
289 ASN 0 37.35 9.62 50.03 15.00
290 LYS N 35.60 9.63 51.45 15.00
290 LYS CA 36.27 10.41 52.48 15.00
290 LYS CB 37.47 9.64 53.05 15.00
290 LYS CG 37.97 10.14 54.42 15.00
290 LYS CD 37.30 9.40 55.60 15.00
290 LYS CE 37.89 9.80 56.97 15.00
290 LYS NZ 37.52 11.19 57.40 15.00
290 LYS C 36.67 11.81 51.97 15.00
290 LYS 0 37.78 12.29 52.23 15.00
291 GLY N 35.78 12.46 51.23 15.00
291 GLY CA 36.06 13.80 50.73 15.00
291 GLY C 36.44 13.95 49.26 15.00
291 GLY 0 36.10 14.95 48.63 15.00
292 TYR N 37.14 12.97 48.70 15.00
292 TYR CA 37.56 13.06 47.31 15.00
292 TYR CB 39.02 12.60 47.16 15.00
292 TYR CG 40.02 13.32 48.04 15.00
292 TYR CDl 40.15 13.01 49.39 15.00
292 TYR CEl 41.03 13.70 50.20 15.00
292 TYR CD2 40.81 14.35 47.53 15.00
292 TYR CE2 41.69 15.04 48.34 15.00
292 TYR CZ 41.79 14.71 49.67 15.00
292 TYR OH 42.66 15.41 50.48 15.00
292 TYR C 36.67 12.18 46.45 15.00
292 TYR 0 35.79 11.46 46.95 15.00
293 ILE N 36.93 12.21 45.15 15.00
293 ILE CA 36.19 11.43 44.19 15.00
293 ILE CB 34.98 12.20 43.68 15.00
293 ILE CG2 35.43 13.55 .43.14 15.00
293 ILE CGI 34.25 11.39 42.62 15.00 293 ILE CDl 33.05 12.09 42.03 15.00
293 ILE C 37.13 11.19 43.02 15.00
293 ILE 0 37.95 12.05 42.70 15.00
294 LEU N 37.05 10.00 42.42 15.00
294 LEU CA 37.88 9.64 41.28 15.00
294 LEU CB 38.29 8.18 41.37 15.00
294 LEU CG 39.48 7.75 42.21 15.00
294 LEU CDl 39.28 8.16 43.66 15.00
294 LEU CD2 39.61 6.25 42.11 15.00
294 LEU C 37.04 9.80 40.04 15.00
294 LEU 0 36.16 9.00 39.80 15.00
295 MET N 37.31 10.81 39.24 15.00
295 MET CA 36.51 11.00 38.03 15.00
295 MET CB 36.25 12.49 37.81 15.00
295 MET CG 35.59 13.13 39.01 15.00
295 MET SD 35.02 14.79 38.73 15.00
295 MET CE 33.94 14.53 37.34 15.00
295 MET C 37.18 10.37 36.83 15.00
295 MET 0 38.41 10.29 36.77 15.00
296 ALA N 36.38 9.92 35.86 15.00
296 ALA CA 36.92 9.29 34.65 15.00
296 ALA CB 35.81 8.97 33.68 15.00
296 ALA C 37.94 10.22 34.01 15.00
296 ALA 0 37.72 11.44 33.93 15.00
297 ARG N 39.04 9.62 33.55 15.00
297 ARG CA 40.14 10.38 32.96 15.00
297 ARG CB 41.48 9.83 33.45 15.00
297 ARG CG 42.65 10.70 33.07 15.00
297 ARG CD 43.59 10.86 34.23 15.00
297 ARG NE 44.34 9.64 34.52 15.00
297 ARG CZ 45.14 9.04 33.64 15.00
297 ARG NHl 45.30 9.53 32.41 15.00
297 ARG NH2 45.85 7.98 34.03 15.00
297 ARG C 40.20 10.55 31.45 15.00
297 ARG 0 39.79 11.60 30.94 15.00
298 ASN N 40.79 9.59 30.75 15.00
298 ASN CA 40.89 9.73 29.30 15.00 298 ASN CB 42.14 9.04 28.77 15.00
298 ASN CG 43.39 9.92 28.86 15.00
298 ASN ODl 43.35 11.11 28.55 15.00
298 ASN ND2 44.50 9.33 29.30 15.00
298 ASN C 39.64 9.27 28.58 15.00
298 ASN 0 39.66 8.32 27.79 15.00
299 LYS N 38.53 9.94 28.89 15.00
299 LYS CA 37.22 9.70 28.31 15.00
299 LYS CB 36.27 9.08 29.34 15.00
299 LYS CG 36.69 7.69 29.84 15.00
299 LYS CD 36.83 6.71 28.68 15.00
299 LYS CE 35.49 6.19 28.19 15.00
299 LYS NZ 34.95 5.23 29.18 15.00
299 LYS C 36.72 11.07 27.91 15.00
299 LYS 0 36.49 11.92 28.76 15.00
300 ASN N 36.63 11.30 26.61 15.00
300 ASN CA 36.21 12.55 26.00 15.00
300 ASN CB 35.05 12.31 25.04 15.00
300 ASN CG 35.52 11.73 23.73 15.00
300 ASN ODl 36.22 10.71 23.71 15.00
300 ASN ND2 35.15 12.38 22.63 15.00
300 ASN C 35.95 13.80 26.83 15.00
300 ASN 0 34.96 14.50 26.62 15.00
301 ASN N 36.88 14.12 27.71 15.00
301 ASN CA 36.77 15.29 28.56 15.00
301 ASN CB 36.73 16.54 27.67 15.00
301 ASN CG 36.90 17.82 28.45 15.00
301 ASN ODl 37.58 17.85 29.49 15.00
301 ASN ND2 36.30 18.90 27.96 15.00
301 ASN C 35.51 15.17 29.41 15.00
301 ASN 0 34.67 16.08 29.47 15.00
302 ALA N 35.37 14.03 30.08 15.00
302 ALA CA 34.21 13.77 30.93 15.00
302 ALA CB 34.41 12.48 31.70 15.00
302 ALA C 33.97 14.93 31.89 15.00
302 ALA 0 34.91 15.48 32.46 15.00
303 CYS N 32.72 15.35 32.01 15.00 303 CYS CA 32.35 16.45 32.89 15.00
303 CYS C 33.20 17.67 32.59 15.00
303 CYS 0 33.24 18.63 33.37 15.00
303 CYS CB 32.55 16.04 34.35 15.00
303 CYS SG 31.58 14.59 34.86 15.00
304 GLY N 33.86 17.64 31.43 15.00
304 GLY CA 34.72 18.72 31.02 15.00
304 GLY C 35.77 19.04 32.05 15.00
304 GLY 0 36.01 20.22 32.32 15.00
305 ILE N 36.43 18.03 32.61 15.00
305 ILE CA 37.44 18.30 33.62 15.00
305 ILE CB 37.95 17.03 34.32 15.00
305 ILE CG2 38.74 16.18 33.37 15.00
305 ILE CGI 38.78 17.42 35.54 15.00
305 ILE CDl 39.35 16.26 36.28 15.00
305 ILE C 38.60 19.14 33.08 15.00
305 ILE 0 39.19 19.95 33.80 15.00
306 ALA N 38.85 19.03 31.79 15.00
306 ALA CA 39.93 19.80 31.19 15.00
306 ALA CB 40.64 18.95 30.18 15.00
306 ALA C 39.42 21.08 30.54 15.00
306 ALA 0 40.01 21.56 29.58 15.00
307 ASN N 38.35 21.67 31.07 15.00
307 ASN CA 37.81 22.89 30.49 15.00
307 ASN CB 36.29 22.91 30.59 15.00
307 ASN CG 35.60 22.56 29.28 15.00
307 ASN ODl 36.23 22.08 28.34 15.00
307 ASN ND2 34.30 22.79 29.23 15.00
307 ASN C 38.35 24.20 31.03 15.00
307 ASN 0 38.76 25.07 30.27 15.00
308 LEU N 38.30 24.36 32.35 15.00
308 LEU CA 38.76 25.60 32.99 15.00
308 LEU CB 37.56 26.32 33.61 15.00
308 LEU CG 36.91 27.48 32.85 15.00
308 LEU CDl 36.75 27.18 31.36 15.00
308 LEU CD2 35.55 27.76 33.51 15.00
308 LEU C 39.79 25.36 34.06 15.00 308 LEU O 39.57 25.69 35.23 15.00
309 ALA N 40.94 24.85 33.64 15.00
309 ALA CA 42.03 24.54 34.57 15.00
309 ALA CB 42.75 23.29 34.11 15.00
309 ALA C 43.01 25.69 34.70 15.00
309 ALA O 43.30 26.36 33.71 15.00
310 SER N 43.56 25.88 35.90 15.00
310 SER CA 44.53 26.93 ■36.16 15.00
310 SER CB 43.85 28.30 36.08 15.00
310 SER OG 42.70 28.40 36.90 15.00
310 SER C 45.23 26.74 37.50 15.00
310 SER O 44.78 25.96 38.34 15.00
311 PHE N 46.36 27.40 37.67 15.00
311 PHE CA 47.12 27.31 38.92 15.00
311 PHE CB 47.96 26.03 38.93 15.00
311 PHE CG 49.01 25.98 37.85 15.00
311 PHE CDl 50.25 26.56 38.04 15.00
311 PHE CD2 48.74 25.37 36.64 15.00
311 PHE CEl 51.21 26.54 37.04 15.00
311 PHE CE2 49.70 25.34 35.64 15.00
311 PHE CZ 50.93 25.93 35.85 15.00
311 PHE C 47.99 28.54 39.03 15.00
311 PHE O 48.41 29.12 •38.02 15.00
312 PRO N 48.25 29.00 40.27 15.00
312 PRO CD 47.72 28.46 41.53 15.00
312 PRO CA 49.07 30.17 40.52 15.00
312 PRO CB 48.67 30.54 41.94 15.00
312 PRO CG 48.52 29.21 42.56 15.00
312 PRO C 50.56 29.86 40.47 15.00
312 PRO O 50.97 28.70 40.58 15.00
313 LYS N 51.37 30.91 40.29 15.00
313 LYS CA 52.82 30.83 40.25 15.00
313 LYS CB 53.38 31.51 39.00 15.00
313 LYS CG 53.01 30.87 37.69 15.00
313 LYS CD 53.59 31.66 36.51 15.00
313 LYS CE 53.12 33.11 36.50 15.00
313 LYS NZ 53.57 33.86 35.30 15.00 313 LYS C 53..26 31..61 41,.46 15,.00
313 LYS O 53. .37 32. .84 41, .41 15, .00
314 MET N 53, .37 30. .92 42. .58 15, .00
314 MET CA 53. .80 31. .54 43, .82 15, .00
314 MET CB 53. .53 30. .56 44, .97 15. .00
314 MET CG 54, .01 31. .02 46. .31 15, .00
314 MET SD 53, .49 29. .88 47, .59 15. .00
314 MET CE 53. .68 30. .97 49, .05 15. .00
314 MET C 55, .29 31, .91 43, .73 15, .00
314 MET OTl 55, .71 32. .88 44, .41 15, .00
314 MET OT2 56. .02 31. .24 42, .96 15. .00
502 HOH OH2 38, .18 18, .74 57, .83 15. .00
503 HOH OH2 41, .01 35. .17 62, .98 15. .00
505 HOH OH2 44. .29 24. .31 '61. .53 15. ,00
509 HOH OH2 45, .95 46, .05 53, .57 15. ,00
510 HOH OH2 30, .32 -1. .58 47, .62 15, .00
511 HOH OH2 37, .81 6, .12 45, .60 15. .00
512 HOH OH2 48, .03 7. .94 27, .32 15, .00
514 HOH OH2 30. .21 15. .76 57, .13 15, .00
515 HOH OH2 45, .27 21. .35 49, .85 15. .00
516 HOH OH2 46, .41 46. .23 50. .69 15, .00
517 HOH OH2 53. .91 41. .75 43. .17 15. ,00
518 HOH OH2 20, .50 21, .52 37. .73 15, .00
519 HOH OH2 21. .73 20. .43 33. .70 15. .00
520 HOH OH2 27, .61 12. .32 27. .20 15, .00
521 HOH OH2 47, .60 46. .15 47. .79 15, .00
522 HOH OH2 61. .85 26. .15 53. .68 15. .00
523 HOH OH2 43, .95 24. .65 64, .43 15. .00
Table II Provides the distances between interresidue atoms that are within 5 Angstroms apart in the active site of the procathepsin
K according to this invention.
Atom 1 Atom 2 Distance Atom 1 Atom 2 Distance
75 N 121 0 4. .995 75 CA 118 NE2 3, .873
75 CA 121 0 4, .355 75 CA 118 CD 4 .497
75 CA 283 NEl 4, .706 75 CA 118 OEl 4 .948
75 CA 119 0 4. .987 75 C 118 NE2 3, .466
75 C 118 CD 4, .018 75 C 118 OEl 4 .145
75 C 283 NEl 4. .154 75 C 121 O 4, .560
75 C 283 CZ2 4, .800 75 C 283 CE2 4 .835
75 C 261 CEl 4, .979 75 0 283 NEl 3 .017
75 0 118 OEl 3 , .398 75 0 118 NE2 3, .416
75 0 118 CD 3, .559 75 0 283 CE2 3. .721
75 0 283 CZ2 3, .785 75 0 261 CEl 3, .966
75 0 283 CDl 4, .133 75 0 261 NDl 4 .354
75 0 118 CG 4, .672 75 0 261 NE2 4, .856
75 CB 283 NEl 3. .971 75 CB 118 NE2 4 .521
75 CB 283 CE2 4, .555 75 CB 119 0 4, .622
75 CB 283 CDl 4, .636 75 CB 118 CD 4, .756
75 CB 283 CZ2 4, .817 75 CG 119 0 3, .992
75 CG 119 C 4, .854 75 CG 283 NEl 4, .957
75 SD 119 0 4 .544 75 SD 283 NEl 4 .903
75 SD 283 CDl 4, .988 75 CE 119 O 3, .405
75 CE 119 C 4 .162 75 CE 283 CDl 4 .953
76 N 118 NE2 3, .957 76 N 121 0 4, .698
76 N 118 CD 4, .726 76 N 118 OEl 4, .792
76 N 122 CA 4 .921 76 CA 261 NDl 4, .016
76 CA 124 CB 4, .124 76 CA 118 NE2 4, .359
76 CA 260 0 4 .376 76 CA 124 SG 4 .392
76 CA 261 CEl 4, .402 76 CA 118 OEl 4, .758
76 CA 118 CD 4, .992 76 C 124 SG 3, .895
76 C 124 CB 3, .957 76 C 122 CA 4, .219
76 C 260 0 4, .588 76 C 118 NE2 4, .643
76 C 122 C 4, .746 76 C 124 N 4, .779
76 C 261 NDl 4, .945 76 0 122 CA 2, .996
76 0 122 C 3. .572 76 O 122 O 3, .992
76 0 124 CB 3, .996 76 O 118 NE2 4, .014
76 0 121 0 4, .109 76 O 124 N 4. .132
76 0 122 N 4, .146 76 O 123 N 4, .207 76 0 124 SG 4.,211 76 0 121 C 4.,541
76 0 124 CA 4. .711 76 CB 260 O 3. .250
76 CB 261 NDl 3. ,886 76 CB 260 C 4. ,410
76 CB 261 CEl 4. .599 76 CB 261 CG 4. ,680
76 CB 261 CA 4. ,690 76 CB 261 CB 4. ,819
76 CB 124 SG 4. ,909 76 OGl 260 O 3, ,483
76 OGl 260 C 4. ,657 76 OGl 260 CB 4, ,921
76 CG2 261 NDl 3. ,842 76 CG2 260 O 3. ,973
76 CG2 283 CZ2 4, .184 76 CG2 261 CEl 4, .378
76 CG2 261 CG 4. ,410 76 CG2 261 CB 4. .672
76 CG2 260 ODl 4, .817 76 CG2 260 C 4, .945
77 N 124 SG 3. .789 77 N 260 O 3. .832
77 N 124 CB 4. .431 77 N 260 C 4, .971
77 N 122 CA 4. .995 77 CA 124 SG 4. .104
77 CA 164 CA 4, .268 77 CA 260 O 4. .740
77 CA 122 CA 4, .753 77 CA 122 O 4. .869
77 CA 164 N 4, .965 77 CA 124 CB 4, .989
77 C 124 SG 3, .712 77 C 164 CA 4. .149
77 C 165 N 4, .290 77 C 165 O 4. .329
77 C 260 0 4, .727 77 C 164 C 4, .784
77 0 165 N 3, .086 77 O 164 CA 3. .304
77 0 165 0 3 .351 77 O 164 C 3, .677
77 0 165 C 4, .096 77 O 124 SG 4. .131
77 0 165 CA .156 77 O 125 CDl 4, .373
77 0 164 N 4 .519 77 O 125 CB 4, .682
77 0 122 0 4 .772 77 O 125 CG 4, .846
77 0 164 0 4 .878 77 O 125 N 4, .998
78 N 124 SG 3, .624 78 N 260 O 3. .772
78 N 260 C 4 .354 78 N 165 O 4. .525
78 N 260 CA 4 .893 78 CA 165 O 3, .675
78 CA 124 SG 4 .038 78 CA 260 O 4, .469
78 CA 260 C 4 .694 78 CA 165 C 4, .858
78 C 165 0 3 .470 78 C 165 C 4 .671
78 C 165 N 4 .812 78 O 165 O 4, .549
78 0 260 CA 4 .953 78 CB 260 C 3, .787
78 CB 260 0 3. .940 78 CB 261 N 4, .054
78 CB 260 CA 4 .128 78 CB 124 SG 4, .281
78 CB 261 C 4 .364 78 CB 261 CA 4 .454
78 CB 261 0 4 .473 78 CB 260 N 4, .727
78 CB 262 N 4 .729 78 CB 165 O 4, .763
78 CG 166 CD2 4 .339 78 CG 233 CB 4 .450
78 CG 165 0 4 .601 78 CG 166 CE2 4. .656
78 CG 261 0 4 .703 78 CG 260 C 4, .930 78 CG 261 C 4,.931 78 CG 261 N 4,.956
78 CDl 167 SD 3. ,767 78 CDl 166 CD2 3. .914
78 CDl 262 CB 4, .020 78 CDl 167 CE 4, .083
78 CDl 233 CB 4, .196 78 CDl 166 CE2 4. .214
78 CDl 165 0 4. ,482 78 CDl 261 0 4. .510
78 CDl 262 CA 4. .616 78 CDl 261 C 4, .720
78 CDl 262 N 4. .747 78 CDl 124 SG 4, .832
78 CDl 166 CA 4. .850 78 CDl 166 CG 4. .853
78 CD2 233 CB 3. .462 78 CD2 261 0 4. .404
78 CD2 260 N 4, .528 78 CD2 233 CA 4, .611
78 CD2 260 CA 4. .717 78 CD2 261 N 4, .737
78 CD2 166 CE2 4. .755 78 CD2 166 CD2 4, .849
78 CD2 260 C 4. .851 78 CD2 261 C 4, .961
79 N 165 0 2. .577 79 N 165 C 3, .666
79 N 165 N 3. .694 79 N 165 CA 4, .214
79 N 164 C 4. .711 79 N 166 N 4, .779
79 N 164 CA 4, .792 79 CA 165 0 3. .276
79 CA 165 N 4. .098 79 CA 165 C 4. .120
79 CA 165 CA 4. .486 79 C 165 0 3. .582
79 C 165 C 4. .428 79 C 166 CD2 4. .589
79 C 166 CB 4, .702 79 0 165 0 3, .305
79 0 166 CB 3. .633 79 0 166 CD2 3. .799
79 0 165 C 3. .919 79 0 166 CG 4. .132
79 0 166 CA 4, .358 79 0 166 N 4, .424
79 0 165 CA 4. .788 79 0 166 CE2 4, .878
79 CB 165 0 3. .121 79 CB 165 N 3. .244
79 CB 165 CA 3. .495 79 CB 165 C 3, .524
79 CB 164 C 4. .048 79 CB 164 CA 4. ,500
79 CB 166 N 4, .607 79 CB 164 0 4, .838
79 CG 165 N 4. .216 79 CG 165 CA 4, .486
79 CG 165 0 4. .575 79 CG 164 C 4. .673
79 CG 165 C 4, .798 79 CD 165 N 4, .409
79 CD 164 CA 4, .436 79 CD 164 C 4. .493
Table III Provides the bond angles between interresidue atoms that are within 5 Angstroms apart in the active site region of procathepsin K according to this invention.
Atom 1 Atom 2 Atom 3 Angle Atom 1 Atom 2 Atom 3 Angle
118NE2 75CA 1210 36, .06 118NE2 75CA 118CD 16, .04
118NE2 75CA 283NE1 66, .79 118NE2 75CA 1180E1 25. .84
118NE2 75CA 1190 58, .31 1210 75CA 118CD 50, .49
1210 75CA 1180E1 61, .89 1210 75CA 1190 58. .01
118CD 75CA 283NE1 51. .41 118CD 75CA 1180E1 13. .90
118CD 75CA 1190 54. .73 283NE1 75CA 1180E1 41. .86
283NE1 75CA 1190 75. .92 1180E1 75CA 1190 66. .13
118NE2 75C 118CD 18. .51 118NE2 75C 1180E1 32. .60
118NE2 75C 283NE1 76. .97 118NE2 75C 1210 34. .31
118NE2 75C 283CE2 91. .87 118NE2 75C 261CE1 75. .88
118CD 75C 1180E1 17. .20 118CD 75C 283NE1 58. .52
118CD 75C 1210 51. .78 118CD 75C 283CZ2 87. .90
118CD 75C 283CE2 73. .37 118CD 75C 261CE1 63. .49
1180E1 75C 283NE1 49. .23 1180E1 75C 1210 66. .91
1180E1 75C 283CZ2 74. .96 1180E1 75C 283CE2 62. .58
1180E1 75C 261CE1 46. .33 283NE1 75C 283CZ2 31. .41
283NE1 75C 283CE2 15. .20 283NE1 75C 261CE1 48. .42
283CZ2 75C 283CE2 16. .61 283CZ2 75C 261CE1 48. ,64
283CE2 75C 261CE1 49. .08 283NE1 750 1180E1 64. .89
283NE1 750 118NE2 95. .58 283NE1 750 118CD 74. .33
283NE1 750 283CE2 20, .15 283NE1 750 283CZ2 41. .26
283NE1 750 261CE1 64. .66 283NE1 750 283CD1 12. .81
283NE1 750 261ND1 79, .83 283NE1 750 118CG 66, .93
283NE1 750 261NE2 53. .74 1180E1 750 118NE2 38. .26
1180E1 750 118CD 20. .14 1180E1 750 283CE2 82, .53
1180E1 750 283CZ2 98. .99 1180E1 750 261CE1 59. .18
1180E1 750 283CD1 57. .60 1180E1 750 261ND1 72, .46
1180E1 750 118CG 29, .49 1180E1 750 261NE2 62, .96
118NE2 750 118CD 21. .71 118NE2 750 261CE1 92. .08
118NE2 750 283CD1 84. .38 118NE2 750 261ND1 99. .59
118NE2 750 118CG 29. .92 118NE2 750 261NE2 99. .17
118CD 750 283CE2 94. .09 118CD 750 261CE1 79. .13
118CD 750 283CD1 63. .81 118CD 750 261ND1 91. .35
118CD 750 118CG 14, .54 118CD 750 261NE2 83. .02
283CE2 750 283CZ2 21. .35 283CE2 750 261CE1 63. ,98 283CE2 750 283CD1 32.,37 283CE2 750 261ND1 75..00
283CE2 750 118CG 87. ,07 283CE2 750 261NE2 51, .36
283CZ2 750 261CE1 62. ,61 283CZ2 750 283CD1 53. .70
283CZ2 750 261ND1 67. .57 283CZ2 750 261NE2 50. .53
261CE1 750 283CD1 71. ,28 261CE1 750 261ND1 17, .46
261CE1 750 118CG 87. ,41 261CE1 750 261NE2 12. .73
283CD1 750 261ND1 87. .81 283CD1 750 118CG 55. .01
283CD1 750 261NE2 61. .92 261ND1 750 261NE2 26, .12
118CG 750 261NE2 88. ,52 283NE1 75CB 118NE2 68. .03
283NE1 75CB 283CE2 16, .72 283NE1 75CB 1190 87. .60
283NE1 75CB 283CD1 15. ,99 283NE1 75CB 118CD 53. .59
283NE1 75CB 283CZ2 31, .23 118NE2 75CB 283CE2 83, .45
118NE2 75CB 1190 57, ,88 118NE2 75CB 283CD1 67, .64
118NE2 75CB 118CD 16. .12 118NE2 75CB 283CZ2 90, .96
283CE2 75CB 283CD1 28. .00 283CE2 75CB 118CD 69, .80
283CE2 75CB 283CZ2 16, .78 1190 75CB 283CD1 73, .70
1190 75CB 118CD 55. .68 283CD1 75CB 118CD 51. .68
283CD1 75CB 283CZ2 44, .58 118CD 75CB 283CZ2 79, .88
1190 75CG 119C 11. .39 1190 75CG 283NE1 82, .86
119C 75CG 283NE1 94. .25 1190 75SD 283NE1 78, .22
1190 75SD 283CD1 71. .07 283NE1 75SD 283CD1 15, .84
1190 75CE 119C 14. .76 1190 75CE 283CD1 80, .96
119C 75CE 283CD1 94. .94 118NE2 76N 1210 33, .42
118NE2 76N 118CD 14, .27 li8NE2 76N 1180E1 27 .52
118NE2 76N 122CA 56, .37 1210 76N 118CD 47, .26
1210 76N 1180E1 60. ,91 1210 76N 122CA 32, .17
118CD 76N 1180E1 14. .79 118CD 76N 122CA 70, .12
1180E1 76N 122CA 79. .60 261ND1 76CA 124CB 54, .84
261ND1 76CA 118NE2 90, .77 261ND1 76CA 260O 61 .33
261ND1 76CA 124SG 58, .56 261ND1 76CA 261CE1 17, .26
261ND1 76CA 1180E1 63 .29 261ND1 76CA 118CD 77 .49
124CB 76CA 118NE2 62, .81 124CB 76CA 260O 85 .35
124CB 76CA 124SG 24, .21 124CB 76CA 261CE1 53, .22
124CB 76CA 1180E1 51, .81 124CB 76CA 118CD 58 .94
118NE2 76CA 124SG 85, .37 118NE2 76CA 261CE1 74, .91
118NE2 76CA 1180E1 27 .92 118NE2 76CA 118CD 14 .28
260O 76CA 124SG 65, .30 260O 76CA 261CE1 78 .56
124SG 76CA 261CE1 64 .93 124SG 76CA 1180E1 75 .85
124SG 76CA 118CD 83 .00 261CE1 76CA 1180E1 47 .04
261CE1 76CA 118CD 61, .13 1180E1 76CA 118CD 14, .20
124SG 76C 124CB 26 .57 124SG 76C 122CA 99 .52
124SG 76C 260O 67, .22 124SG 76C 118NE2 87 .57
124SG 76C 122C 81 .40 124SG 76C 124N 48 .42 124SG 76C 261ND1 54,.07 124CB 76C 122CA 87..74
124CB 76C 260O 84, .53 124CB 76C 118NE2 61, .31
124CB 76C 122C 70, .61 124CB 76C 124N 31, .25
124CB 76C 261ND1 48, .28 122CA 76C 118NE2 57, .49
122CA 76C 122C 18, .19 122CA 76C 124N 56, .49
260O 76C 261ND1 53. .35 118NE2 76C 122C 55, .01
118NE2 76C 124N 47. .65 118NE2 76C 261ND1 76. .88
122C 76C 124N 39. .55 124N 76C 261ND1 77, .02
122CA 760 122C 24. .62 122CA 760 1220 36, .26
122CA 760 118NE2 73. .62 122CA 760 1210 40, .55
122CA 760 124N 71. .96 122CA 760 122N 14, .30
122CA 760 123N 34, .47 122CA 760 121C 28, .56
122CA 760 124CA 89, .43 122C 760 1220 17, .57
122C 760 124CB 83, .93 122C 760 118NE2 69, .46
122C 760 1210 48, .92 122C 760 124N 48, .79
122C 760 122N 36, .55 122C 760 123N 17, .28
122C 760 124SG 93, .20 122C 760 121C 44, .07
122C 760 124CA 65, .97 1220 760 124CB 80, .66
1220 760 118NE2 84, .24 1220 760 1210 66, .48
1220 760 124N 49, .34 1220 760 122N 50, .21
1220 760 123N 31, .60 1220 760 124SG 83, .04
1220 760 121C 60, .61 1220 760 124CA 64, .22
124CB 760 118NE2 67, .06 124CB 760 1210 96, .67
124CB 760 124N 35, .52 124CB 760 123N 73, .71
124CB 760 124SG 25, .24 124CB 760 124CA 18. .06
118NE2 760 1210 37, .13 118NE2 760 124N 55, .72
118NE2 760 122N 67, .51 118NE2 760 123N 52. .88
118NE2 760 124SG 92. .30 118NE2 760 121C 52. .23
118NE2 760 124CA 60, .34 1210 760 124N 69, .55
1210 760 122N 31, .11 1210 760 123N 40, .00
1210 760 121C 15, .12 1210 760 124CA 83. .05
124N 760 122N 79, .70 124N 760 123N 38. .53
124N 760 124SG 51, .84 124N 760 121C 76. .98
124N 760 124CA 17, .47 122N 760 123N 41. .20
122N 760 121C 16, .68 122N 760 124CA 96. .89
123N 760 124SG 88, .96 123N 760 121C 41. .29
123N 760 124CA 55, .85 124SG 760 124CA 36. .87
121C 760 124CA 92. .58 2600 76CB 261ND1 73. .33
260O 76CB 260C 5, .97 2600 76CB 261CE1 88. .42
260O 76CB 261CG 64. .24 2600 76CB 261CA 33, .95
260O 76CB 261CB 47. .27 2600 76CB 124SG 67. ,36
261ND1 76CB 260C 69. .61 261ND1 76CB 261CE1 15, .10
261ND1 76CB 261CG 15. .13 261ND1 76CB 261CA 39. .38 261ND1 76CB 261CB 31..52 261ND1 76CB 124SG 54.44
260C 76CB 261CE1 84. .69 260C 76CB 26ICG 59 .58
260C 76CB 261CA 30, .41 260C 76CB 261CB 42 .17
260C 76CB 124SG 69. .37 261CE1 76CB 261CG 27 .44
261CE1 76CB 261CA 54. .48 261CE1 76CB 261CB 45 .34
261CE1 76CB 124SG 59. .43 26ICG 76CB 261CA 31 .56
261CG 76CB 261CB 18. .14 26ICG 76CB 124SG 63, .87
261CA 76CB 261CB 18, .49 261CA 76CB 124SG 54 .00
261CB 76CB 124SG 66, .72 260O 760G1 260C 5 .04
260O 760G1 260CB 34. .36 260C 760G1 260CB 29, .73
261ND1 76CG2 260O 66. .59 261ND1 76CG2 283CZ2 68, .93
261ND1 76CG2 261CE1 16, .92 261ND1 76CG2 26ICG 17 .51
261ND1 76CG2 261CB 32, .75 261ND1 76CG2 260OD1 81 .35
261ND1 76CG2 260C 64. .09 260O 76CG2 261CE1 83, .30
260O 76CG2 261CG 62. .86 260O 76CG2 261CB 47, .50
260O 76CG2 260OD1 42. .82 260O 76CG2 260C 9, .65
283CZ2 76CG2 261CE1 56, .11 283CZ2 76CG2 261CG 66, .23
283CZ2 76CG2 261CB 80, .03 261CE1 76CG2 261CG 29, .03
261CE1 76CG2 261CB 47. .23 261CE1 76CG2 260OD1 95, .65
261CE1 76CG2 260C 81. .00 261CG 76CG2 261CB 18, .77
261CG 76CG2 260OD1 66, .64 261CG 76CG2 260C 57, .48
261CB 76CG2 260OD1 48. .72 261CB 76CG2 260C 40, .55
260OD1 76CG2 260C 34. .76 124SG 77N 260O 76, .74
124SG 77N 124CB 23. .76 124SG 77N 260C 73. .38
124SG 77N 122CA 88, .63 260O 77N 124CB 88. .17
260O 77N 260C 5, .96 124CB 77N 260C 86, .75
124CB 77N 122CA 73, .67 124SG 77CA 260O 64, .21
124SG 77CA 122CA 88, .49 124SG 77CA 1220 74. .09
124SG 77CA 124CB 20, ,03 164CA 77CA 122CA 70. .18
164CA 77CA 1220 56. .41 164CA 77CA 164N 15, .89
260O 77CA 124CB 72, .64 122CA 77CA 1220 28. .50
122CA 77CA 164N 57, .26 122CA 77CA 124CB 71. .13
1220 77CA 164N 50. ,18 1220 77CA 124CB 63, ,25
124SG 77C 1650 94, .31 124SG 77C 260O 66. .95
164CA 77C 165N 32, .95 164CA 77C 1650 69. .26
164CA 77C 164C 17, .70 165N 77C 1650 36, ,54
165N 77C 164C 15. ,64 1650 77C 164C 52. ,18
165N 770 164CA 43. ,90 165N 770 1650 49. ,43
165N 770 164C 20, .33 165N 770 165C 36, .57
165N 770 165CA 15, ,71 165N 770 125CD1 57, ,18
165N 770 164N 53. ,60 165N 770 125CB 67. ,06
165N 770 1220 78. .73 165N 770 125CG 56. ,08
165N 770 1640 20, .42 165N 770 125N 92, .35 164CA 770 1650 92,.82 164CA 770 164C 24,.23
164CA 770 165C 80, .47 164CA 770 165CA 59 .42
164CA 770 125CD1 69, .56 164CA 770 164N 11 .76
164CA 770 125CB 97. .27 164CA 770 1220 62, .09
164CA 770 125CG 79, .85 164CA 770 1640 26 .06
1650 770 164C 69, .76 1650 770 165C 15 .26
1650 770 165CA 35, .08 1650 770 125CD1 73, .77
1650 770 125CB 53, .64 1650 770 125CG 60, .06
1650 770 1640 69, .49 1650 770 125N 81 .29
164C 770 165C 56, .55 164C 770 165CA 35, .40
164C 770 125CD1 56, .97 164C 770 164N 33, .30
164C 770 125CB 77 .67 164C 770 1220 66 .26
164C 770 125CG 62, .33 164C 770 1640 3, .99
164C 770 125N 96, .65 165C 770 165CA 21, .15
165C 770 125CD1 61, .03 165C 770 164N 89, .72
165C 770 125CB 47, .59 165C 770 1220 96, .34
165C 770 125CG 49, .12 165C 770 1640 55, .72
165C 770 125N 76, .96 124SG 770 125CD1 77, .46
124SG 770 125CB 64, .58 124SG 770 1220 74, .93
124SG 770 125CG 75, .13 124SG 770 125N 38, .93
165CA 770 125CD1 54, .72 165CA 770 164N 68. .58
165CA 770 125CB 55, .56 165CA 770 1220 84. .07
165CA 770 125CG 48, .67 165CA 770 1640 34, .63
165CA 770 125N 83, .50 125CD1 770 164N 67, .76
125CD1 770 125CB 32, .79 125CD1 770 1220 37. .02
125CD1 770 125CG 15, .92 125CD1 770 1640 52. .99
125CD1 770 125N 39, .98 164N 770 125CB 98. .50
164N 770 1220 53. .25 164N 770 125CG 80. .38
164N 770 1640 34, .03 125CB 770 1220 65, .89
125CB 770 125CG 18, .18 125CB 770 1640 74. .21
125CB 770 125N 29, .43 1220 770 125CG 52. .30
1220 770 1640 63. .19 1220 770 125N 54, .39
125CG 770 1640 58, .58 125CG 770 125N 36, .28
1640 770 125N 92. .69 124SG 78N 260O 79. ,51
124SG 78N 260C 83. .08 124SG 78N 1650 92, .32
2600 78N 260C 15, .29 260O 78N 26OCA 28. .16
260C 78N 26OCA 17, .65 1650 78CA 124SG 99. .93
1650 78CA 165C 4. .78 124SG 78CA 260O 67, .35
124SG 78CA 260C 74. .63 124SG 78CA 165C 95. ,67
260O 78CA 260C 15, .11 1650 78C 165C 4, ,04
1650 78C 165N 33. .36 165C 78C 165N 29, .89
260C 78CB 260O 18. .11 260C 78CB 261N 19. ,07
260C 78CB 26OCA 21, .52 260C 78CB 124SG 82, .24 260C 78CB 261C 50..82 260C 78CB 261CA 32.63
260C 78CB 2610 55. .55 260C 78CB 260N 32 .43
260C 78CB 262N 63, .04 260O 78CB 261N 32 .35
260O 78CB 26OCA 34. .09 260O 78CB 124SG 70 .13
260O 78CB 261C 56. .71 260O 78CB 261CA 36 .83
260O 78CB 2610 66, .17 260O 78CB 260N 48 .80
260O 78CB 262N 64. .33 261N 78CB 26OCA 34 .77
261N 78CB 124SG 77, .85 261N 78CB 261C 33 .28
261N 78CB 261CA 18, .95 261N 78CB 2610 36 .48
261N 78CB 260N 36. .74 261N 78CB 262N 47, .56
26OCA 78CB 261C 67, .94 26OCA 78CB 261CA 52 .32
26OCA 78CB 2610 67. .73 26OCA 78CB 260N 17 .14
26OCA 78CB 262N 82. .21 124SG 78CB 261C 58, .84
124SG 78CB 261CA 59, .89 124SG 78CB 2610 73, .12
124SG 78CB 262N 44. .93 124SG 78CB 1650 81, .43
261C 78CB 261CA 19, .89 261C 78CB 2610 15, .94
261C 78CB 260N 66. .21 261C 78CB 262N 16, .25
261CA 78CB 2610 31. .13 261CA 78CB 260N 55, .67
261CA 78CB 262N 30, .41 2610 78CB 260N 61, .05
2610 78CB 262N 28. .30 260N 78CB 262N 82, .18
166CD2 78CG 233CB 92, .45 166CD2 78CG 1650 59, .87
166CD2 78CG 166CE2 17. .29 233CB 78CG 166CE2 76, .12
233CB 78CG 2610 46, .52 233CB 78CG 260C 84, .76
233CB 78CG 261C 60. .56 233CB 78CG 261N 71, .12
1650 78CG 166CE2 77, .08 2610 78CG 260C 47. .65
2610 78CG 261C 14. .42 2610 78CG 261N 32, .31
260C 78CG 261C 42. .03 260C 78CG 261N 15. .40
261C 78CG 261N 28. .44 167SD 78CD1 166CD2 75. .55
167SD 78CD1 262CB 57. .53 167SD 78CD1 167CE 25, .64
167SD 78CD1 233CB 89, .72 167SD 78CD1 166CE2 73. .72
167SD 78CD1 1650 93. ,53 167SD 78CD1 2610 98, ,39
167SD 78CD1 262CA 68, .67 167SD 78CD1 261C 98. .69
167SD 78CD1 262N 85. .99 167SD 78CD1 124SG 90. .32
167SD 78CD1 166CA 64, .21 167SD 78CD1 166CG 67. .34
166CD2 78CD1 167CE 65. ,10 166CD2 78CD1 166CE2 19. .20
166CD2 78CD1 1650 63. ,87 166CD2 78CD1 166CA 42. ,23
166CD2 78CD1 166CG 13, .29 262CB 78CD1 167CE 74, .75
262CB 78CD1 233CB 86. ,41 262CB 78CD1 2610 54, ,29
262CB 78CD1 262CA 18, ,46 262CB 78CD1 261C 46, ,32
262CB 78CD1 262N 30, ,66 262CB 78CD1 124SG 43, .36
167CE 78CD1 233CB 70. .78 167CE 78CD1 166CE2 55. .94
167CE 78CD1 2610 97. ,46 167CE 78CD1 262CA 79. .31
167CE 78CD1 262N 96. ,89 167CE 78CD1 166CA 73. .32 167CE 78CD1 166CG 62,.41 233CB 78CD1 166CE2 83,.80
233CB 78CD1 2610 49. .01 233CB 78CD1 262CA 70 .73
233CB 78CD1 261C 64. .04 233CB 78CD1 262N 73, .78
166CE2 78CD1 1650 83, .05 166CE2 78CD1 166CA 59, .39
166CE2 78CD1 166CG 29. .73 1650 78CD1 124SG 78 .68
1650 78CD1 166CA 34. .26 1650 78CD1 166CG 56, .08
2610 78CD1 262CA 36. .15 2610 78CD1 261C 15, .09
2610 78CD1 262N 28. .16 2610 78CD1 124SG 67, .79
262CA 78CD1 261C 30, .54 262CA 78CD1 262N 17 .84
262CA 78CD1 124SG 50, .40 261C 78CD1 262N 16, .21
261C 78CD1 124SG 52. .78 262N 78CD1 124SG 42, .44
124SG 78CD1 166CA 96, .43 166CA 78CD1 166CG 29 .75
233CB 78CD2 2610 53, .21 233CB 78CD2 260N 92, .15
233CB 78CD2 233CA 14, .15 233CB 78CD2 261N 82, .42
233CB 78CD2 166CE2 84. .68 233CB 78CD2 166CD2 98, .20
233CB 78CD2 260C 97, .86 233CB 78CD2 261C 65, .86
2610 78CD2 260N 63, .15 2610 78CD2 233CA 39, .06
2610 78CD2 26OCA 63. .42 2610 78CD2 261N 34, .09
2610 78CD2 260C 49. .52 2610 78CD2 261C 13, .47
260N 78CD2 233CA 83, .98 260N 78CD2 26OCA 17, .85
260N 78CD2 261N 35. .61 260N 78CD2 260C 31, .22
260N 78CD2 261C 63. .05 233CA 78CD2 26OCA 93. .30
233CA 78CD2 261N 69, .20 233CA 78CD2 166CE2 94, .77
233CA 78CD2 260C 84, .90 233CA 78CD2 261C 51, .75
26OCA 78CD2 261N 29. .99 26OCA 78CD2 260C 18. .16
26OCA 78CD2 261C 58, .71 261N 78CD2 260C 15, .83
261N 78CD2 261C 28, .89 166CE2 78CD2 166CD2 16, .58
260C 78CD2 261C 42, .24 1650 79N 165C 10. .81
1650 79N 165N 47, .00 1650 79N 165CA 30, .61
1650 79N 164C 58. .72 1650 79N 166N 7, .73
1650 79N 164CA 75. .06 165C 79N 165N 38. .87
165C 79N 165CA 20. .71 165C 79N 164C 50. ,56
165C 79N 166N 9, .97 165C 79N 164CA 67. .94
165N 79N 165CA 19. .76 165N 79N 164C 11. .75
165N 79N 166N 48. .84 165N 79N 164CA 29, .34
165CA 79N 164C 30, .89 165CA 79N 166N 30. .45
165CA 79N 164CA 48, .99 164C 79N 166N 60. .54
164C 79N 164CA 18. .26 166N 79N 164CA 77. ,85
1650 79CA 165N 41. .13 1650 79CA 165C 14. .08
1650 79CA 165CA 31. .17 165N 79CA 165C 34. ,67
165N 79CA 165CA 18. .76 165C 79CA 165CA 19. .70
1650 79C 165C 12, .94 1650 79C 166CD2 64. ,92
1650 79C 166CB 46. .02 165C 79C 166CD2 65. .90 165C 79C 166CB 40..97 166CD2 79C 166CB 31,.09
1650 790 166CB 58. .43 1650 790 166CD2 77, .59
1650 790 165C 17. .10 1650 790 166CG 71, .57
1650 790 166CA 39. .48 1650 790 166N 29, .63
1650 790 165CA 27, .24 1650 790 166CE2 87 .44
166CB 790 166CD2 39, .11 166CB 790 165C 50, .05
166CB 790 166CG 20, .86 166CB 790 166CA 19, .18
166CB 790 166N 33. .28 166CB 790 165CA 62, .88
166CB 790 166CE2 50. .06 166CD2 790 165C 78, .93
166CD2 790 166CG 19. .43 166CD2 790 166CA 46, .73
166CD2 790 166N 65. .76 166CD2 790 165CA 95, .01
166CD2 790 166CE2 11. .63 165C 790 166CG 67, .58
165C 790 166CA 33, .62 165C 790 166N 16, .99
165C 790 165CA 16, .49 165C 790 166CE2 90, .16
166CG 790 166CA 33, .98 166CG 790 166N 51, .93
166CG 790 165CA 82, .12 166CG 790 166CE2 29, .53
166CA 790 166N 19, .03 166CA 790 165CA 48, .76
166CA 790 166CE2 58. .34 166N 790 165CA 30. .19
166N 790 166CE2 77. .37 1650 79CB 165N 50. .20
1650 79CB 165CA 41, .86 1650 79CB 165C 20, .25
1650 79CB 164C 66, .94 1650 79CB 164CA 76, .08
1650 79CB 166N 25, .81 1650 79CB 1640 71, .51
165N 79CB 165CA 24, .50 165N 79CB 165C 42. .17
165N 79CB 164C 16, .78 165N 79CB 164CA 31. ,00
165N 79CB 166N 51, .76 165N 79CB 1640 23, .32
165CA 79CB 165C 24, .94 165CA 79CB 164C 36. .59
165CA 79CB 164CA 54, .55 165CA 79CB 166N 31. .14
165CA 79CB 1640 34, .36 165C 79CB 164C 58. .02
165C 79CB 164CA 72. .65 165C 79CB 166N 10, .99
165C 79CB 1640 58, .61 164C 79CB 164CA 19. .44
164C 79CB 166N 66, .75 164C 79CB 1640 12. .33
164CA 79CB 166N 82, .64 164CA 79CB 1640 29. .49
166N 79CB 1640 65. .48 165N 79CG 165CA 18, ,86
165N 79CG 1650 35. .50 165N 79CG 164C 16. ,15
165N 79CG 165C 30. .67 165CA 79CG 1650 30. .56
165CA 79CG 164C 30, .62 165CA 79CG 165C 18. .40
1650 79CG 164C 51. .63 1650 79CG 165C 14, .88
164C 79CG 165C 45. .97 165N 79CD 164CA 31. ,44
165N 79CD 164C 17. .13 164CA 79CD 164C 19. ,46 Table IV Table of the orthogonal three dimensional coordinates in Angstroms and B factors (A^) for procathepsin K active site atoms and atoms of the propeptide in the region of the active site .
Residue Atom X Y Z B
75 MET N 25. .19 25, .27 46 .93 15, .00
75 MET CA 26. .49 24, .70 47. .29 15, .00
75 MET C 27, .52 24. .57 '46, .17 15. .00
75 MET 0 28. .31 23, .63 46. .18 15, .00
75 MET CB 26. .30 23, .34 47, .96 15, .00
75 MET CG 25. .56 23, .38 49, .25 15. .00
75 MET SD 25, .38 21, .74 49 .87 15, .00
75 MET CE 26. .23 21. .91 51, .38 15. .00
76 THR N 27. .50 25 .45 45 .19 15, .00
76 THR CA 28. .48 25, .45 44, .12 15, .00
76 THR C 28. .74 26, .93 43, .86 15. .00
76 THR 0 28, .92 27 .68 44 .82 15, .00
76 THR CB 27. .97 24, .75 42, .84 15. .00
76 THR OGl 26. .90 25, .50 42, .28 15, ,00
76 THR CG2 27. .50 23, .34 43, .14 15. .00
77 GLY N 28. .75 27, .36 42, .60 15, .00
77 GLY CA 28. .96 28 .77 42 .33 15, .00
77 GLY C 30. .07 29, .15 '41, .36 15. .00
77 GLY 0 30. .68 30, .20 41, .51 15. .00
78 LEU N 30. .32 28 .33 40 .35 15. .00
78 LEU CA 31. .38 28, .66 39, .42 15. .00
78 LEU C 30. .86 29, .69 38, .47 15. .00
78 LEU 0 30, .29 29 .33 37 .44 15. .00
78 LEU CB 31. .85 27, .45 38, .61 15. .00
78 LEU CG 33. .00 27 .84 37 .67 15. .00
78 LEU CDl 34. .30 27 .78 38, .44 15. .00
78 LEU CD2 33. .07 26, .95 36, .46 15. ,00
79 LYS N 31. .00 30 .96 38, .81 15. .00
79 LYS CA 30. .57 32, .02 37, .93 15. .00
79 LYS C 31. .52 31. .90 36, .75 15. .00 79 LYS 0 32.58 32.52 .36.75 15.00
79 LYS CB 30.77 33.37 38.60 15.00
79 LYS CG 29.87 34.48 38.12 15.00
79 LYS CD 28.54 34.41 38.82 15.00
79 LYS CE 27.78 35.74 38.71 15.00
79 LYS NZ 28.35 36.82 39.58 15.00
118 GLN N 32.65 23.81 52.55 15.00
118 GLN CA 32.21 25.12 52.08 15.00
118 GLN C 30.86 25.59 52.59 15.00
118 GLN 0 30.32 26.57 52.09 15.00
118 GLN CB 32.17 25.14 50.55 15.00
118 GLN CG 30.98 24.41 49.94 15.00
118 GLN CD 30.83 24.71 48.46 15.00
118 GLN OEl 31.38 24.01 47.61 15.00
118 GLN NE2 30.11 25.77 48.14 15.00
119 GLY N 30.32 24.91 .53.58 15.00
119 GLY CA 29.02 25.31 54.10 15.00
119 GLY C 28.03 25.31 52.96 15.00
119 GLY 0 28.21 24.61 51.97 15.00
121 CYS N 27.33 27.97 51.04 15.00
121 CYS CA 27.63 29.26 50.43 15.00
121 CYS C 28.38 29.01 49.13 15.00
121 CYS 0 28.91 27.93 48.92 15.00
121 CYS CB 28.43 30.13 51.42 15.00
121 CYS SG 29.68 31.27 50.76 15.00
122 GLY N 28.41 29.98 48.24 15.00
122 GLY CA 29.08 29.76 46.98 15.00
122 GLY C 30.58 29.94 47.04 15.00
122 GLY 0 31.13 30.69 46.24 15.00
123 SER N 31.24 29.25 47.96 15.00
123 SER CA 32.69 29.37 •48.08 15.00
123 SER C 33.44 28.36 47.23 15.00
123 SER 0 34.63 28.16 47.44 15.00
123 SER CB 33.10 29.19 49.53 15.00
123 SER OG 32.79 30.33 50.30 15.00
124 CYS N 32.79 27.73 46.27 15.00
124 CYS CA 33.49 26.74 45.45 15.00
124 CYS C 34.71 27.36 44.77 15.00 124 CYS 0 35.80 26.79 44.79 15.00
124 CYS CB 32.55 26.04 44.46 15.00
124 CYS SG 32.47 26.70 42.78 15.00
125 TRP N 34.54 28.58 44.25 15.00
125 TRP CA 35.62 29.31 43.58 15.00
125 TRP C 36.72 29.59 "44.60 15.00
125 TRP 0 37.89 29.34 44.34 15.00
125 TRP CB 35.11 30.63 42.97 15.00
125 TRP CG 34.60 31.64 43.98 15.00
125 TRP CDl 33.40 31.61 44.63 15.00
125 TRP CD2 35.30 32.77 44.49 15.00
125 TRP NEl 33.33 32.64 45.53 15.00
125 TRP CE2 34.48 33.37 45.47 15.00
125 TRP CE3 36.56 33.34 44.23 15.00
125 TRP CZ2 34.87 34.51 46.18 15.00
125 TRP CZ3 36.95 34.47 44.94 15.00
125 TRP CH2 36.10 35.04 45.90 15.00
126 ALA N 36.33 30.05 45.79 15.00
126 ALA CA 37.30 30.34 46.83 15.00
126 ALA C 37.90 29.09 47.45 15.00
126 ALA 0 38.80 29.19 '48.28 15.00
126 ALA CB 36.68 31.21 47.90 15.00
127 PHE N 37.39 27.92 47.12 15.00
127 PHE CA 37.98 26.72 47.69 15.00
127 PHE C 38.98 26.09 46.74 15.00
127 PHE 0 40.05 25.67 47.16 15.00
127 PHE CB 36.92 25.73 48.16 15.00
127 PHE CG 36.48 25.95 49.59 15.00
127 PHE CDl 35.54 26.93 49.90 15.00
127 PHE CD2 37.03 25.21 50.62 15.00
127 PHE CEl 35.17 27.16 51.21 15.00
127 PHE CE2 36.66 25.45 51.95 15.00
127 PHE CZ 35.72 26.42 52.24 15.00
128 SER N 38.66 26.08 45.46 15.00
128 SER CA 39.56 25.54 44.45 15.00
128 SER C 40.85 26.35 '44.50 15.00
128 SER 0 41.95 25.80 44.63 15.00
128 SER CB 38.91 25.64 43.07 15.00 128 SER OG 39.60 24.88 42.09 15.00
129 SER N 40.72 27.68 44.44 15.00
129 SER CA 41.86 28.57 44.47 15.00
129 SER C 42.69 28.33 45.69 15.00
129 SER O 43.87 28.02 45.59 15.00
129 SER CB 41.40 30.01 44.43 15.00
129 SER OG 40.65 30.23 43.25 15.00
130 VAL N 42.07 28.41 46.85 15.00
130 VAL CA 42.79 28.18 48.10 15.00
130 VAL C 43.58 26.88 •48.04 15.00
130 VAL O 44.65 26.79 48.62 15.00
130 VAL CB 41.81 28.14 49.28 15.00
130 VAL CGI 42.27 27.18 50.35 15.00
130 VAL CG2 41.70 29.52 49.85 15.00
131 GLY N 43.08 25.91 47.29 15.00
131 GLY CA 43.74 24.61 47.18 15.00
131 GLY C 45.07 24.66 46.45 15.00
131 GLY O 46.06 24.12 46.94 15.00
149 PRO N 39.82 30.57 52.85 15.00
149 PRO CA 38.92 30.56 51.69 15.00
149 PRO C 38.04 31.80 51.73 15.00
149 PRO O 37.68 32.37 50.70 15.00
149 PRO CB 38.09 29.30 51.91 15.00
149 PRO CG 38.90 28.48 52.90 15.00
149 PRO CD 39.42 29.53 '53.81 15.00
153 VAL N 36.59 35.19 49.34 15.00
153 VAL CA 35.16 35.50 49.32 15.00
153 VAL C 34.78 36.95 49.62 15.00
153 VAL O 34.07 37.57 48.82 15.00
153 VAL CB 34.35 34.61 50.31 15.00
153 VAL CGI 32.85 34.96 50.23 15.00
153 VAL CG2 34.56 33.13 50.03 15.00
161 GLY N 31.35 36.59 45.80 15.00
161 GLY CA 31.46 35.98 47.11 15.00
161 GLY C 30.58 34.75 47.20 15.00
161 GLY O 30.57 33.93 46.28 15.00
162 CYS N 29.77 34.67 48.25 15.00
162 CYS CA 28.88 33.53 48.42 15.00 162 CYS C 27.91 33.35 47.27 15.00
162 CYS 0 27.27 32.31 47.16 15.00
162 CYS CB 28.11 33.63 49.74 15.00
162 CYS SG 29.06 33.14 51.22 15.00
164 GLY N 28.81 33.41 44.09 15.00
164 GLY CA 29.51 32.96 42.90 15.00
164 GLY C 30.80 33.70 42.64 15.00
164 GLY 0 31.08 34.75 43.23 15.00
165 GLY N 31.60 33.14 41.73 15.00
165 GLY CA 32.87 33.73 41.37 15.00
165 GLY C 33.69 32.80 40.50 15.00
165 GLY 0 33.15 31.84 39.94 15.00
166 TYR N 34.99 33.03 40.42 15.00
166 TYR CA 35.87 32.22 39.60 15.00
166 TYR C 37.28 32.18 '40.15 15.00
166 TYR 0 37.67 33.05 40.93 15.00
166 TYR CB 35.91 32.77 38.19 15.00
166 TYR CG 36.63 31.87 37.25 15.00
166 TYR CDl 37.97 32.08 36.95 15.00
166 TYR CD2 35.98 30.82 36.64 15.00
166 TYR CEl 38.64 31.27 36.05 15.00
166 TYR CE2 36.64 30.00 35.73 15.00
166 TYR CZ 37.96 30.23 35.43 15.00
166 TYR OH 38.59 29.45 34.50 15.00
167 MET N 38.08 31.24 39.66 15.00
167 MET CA 39.44 31.09 40.16 15.00
167 MET C 40.51 32.11 39.77 15.00
167 MET 0 41.15 32.69 40.66 15.00
167 MET CB 39.93 29.66 39.91 15.00
167 MET CG 39.21 28.59 40.73 15.00
167 MET SD 37.76 27.96 39.92 15.00
167 MET CE 38.38 27.66 38.30 15.00
188 TYR N 36.56 30.81 58.36 15.00
188 TYR CA 35.52 30.72 57.34 15.00
188 TYR C 34.23 30.68 58.14 15.00
188 TYR 0 34.13 29.94 59.12 15.00
188 TYR CB 35.65 29.41 56.55 15.00
188 TYR CG 34.66 29.26 55.41 15.00 188 TYR CDl 34.74 30.08 54.27 15.00
188 TYR CD2 33.66 28.28 55.45 15.00
188 TYR CEl 33.85 29.92 53.20 15.00
188 TYR CE2 32.77 28.12 54.38 15.00
188 TYR CZ 32.88 28.94 '53.26 15.00
188 TYR OH 32.03 28.77 52.21 15.00
231 SER N 43.74 23.58 39.76 15.00
231 SER CA 42.33 23.89 39.96 15.00
231 SER C 41.54 23.56 38.69 15.00
231 SER 0 42.02 23.82 37.58 15.00
231 SER CB 42.17 25.38 40.29 15.00
231 SER OG 42.60 25.66 41.61 15.00
232 VAL N 40.35 22.96 38.84 15.00
232 VAL CA 39.50 22.62 37.69 15.00
232 VAL C 38.01 22.93 37.92 15.00
232 VAL 0 37.59 23.17 39.06 15.00
232 VAL CB 39.65 21.13 37.25 15.00
232 VAL CGI 41.00 20.91 36.59 15.00
232 VAL CG2 39.47 20.20 38.43 15.00
233 ALA N 37.24 22.97 36.84 15.00
233 ALA CA 35.81 23.26 36.88 15.00
233 ALA C 35.12 22.07 36.25 15.00
233 ALA 0 35.63 21.51 35.29 15.00
233 ALA CB 35.51 24.52 36.09 15.00
234 ILE N 33.95 21.70 36.77 15.00
234 ILE CA 33.20 20.54 36.28 15.00
234 ILE C 31.69 20.74 36.29 15.00
234 ILE 0 31.19 21.79 36.70 15.00
234 ILE CB 33.48 19.29 37.16 15.00
234 ILE CGI 33.34 19.68 38.65 15.00
234 ILE CG2 34.83 18.69 36.84 15.00
234 ILE CDl 33.22 18.53 39.59 15.00
260 ASN N 29.75 23.87 36.36 15.00
260 ASN CA 29.17 24.51 37.52 15.00
260 ASN C 29.82 24.27 38.87 15.00
260 ASN 0 29.30 24.73 39.88 15.00
260 ASN CB 27.70 24.15 37.61 15.00
260 ASN CG 27.47 22.66 37.68 15.00 260 ASN ODl 28.02 21.99 38.54 15.00
260 ASN ND2 26.68 22.14 36.75 15.00
261 HIS N 30.91 23.52 38.92 15.00
261 HIS CA 31.57 23.30 40.20 15.00
261 HIS C 33.07 23.51 40.04 15.00
261 HIS 0 33.61 23.35 38.95 15.00
261 HIS CB 31.25 21.91 40.75 15.00
261 HIS CG 31.51 21.75 42.23 15.00
261 HIS NDl 31.30 22.77 .43.13 15.00
261 HIS CD2 31.94 20.69 42.95 15.00
261 HIS CEl 31.59 22.34 44.35 15.00
261 HIS NE2 31.98 21.08 44.26 15.00
262 ALA N 33.72 23.92 41.14 15.00
262 ALA CA 35.15 24.16 41.18 15.00
262 ALA C 35.81 23.18 42.16 15.00
262 ALA 0 35.49 23.18 43.35 15.00
262 ALA CB 35.41 25.58 41.62 15.00
263 VAL N 36.75 22.39 41.66 15.00
263 VAL CA 37.44 21.39 42.46 15.00
263 VAL C 38.97 21.34 42.25 15.00
263 VAL 0 39.51 21.84 41.26 15.00
263 VAL CB 36.85 19.99 42.18 15.00
263 VAL CGI 35.47 19.89 42.73 15.00
263 VAL CG2 36.82 19.71 .40.69 15.00
264 LEU N 39.66 20.70 43.18 15.00
264 LEU CA 41.11 20.58 43.16 15.00
264 LEU C 41.59 19.25 42.56 15.00
264 LEU 0 41.33 18.20 43.14 15.00
264 LEU CB 41.65 20.69 44.59 15.00
264 LEU CG 43.12 21.07 44.77 15.00
264 LEU CDl 43.26 22.53 44.44 15.00
264 LEU CD2 43.59 20.81 46.19 15.00
281 ASN N 36.94 17.76 46.74 15.00
281 ASN CA 35.74 18.57 46.76 15.00
281 ASN C 35.73 19.48 47.98 15.00
281 ASN 0 36.70 19.50 48.74 15.00
281 ASN CB 34.48 17.70 46.73 15.00
281 ASN CG 33.38 18.32 45.91 15.00 281 ASN ODl 32.82 19.36 46.27 15.00
281 ASN ND2 33.09 17.72 44.77 15.00
282 SER N 34.62 20.20 48.15 15.00
282 SER CA 34.44 21.13 49.26 15.00
282 SER C 33.19 20.80 50.05 15.00
282 SER 0 32.67 21.65 50.76 15.00
282 SER CB 34.30 22.55 48.70 15.00
282 SER OG 34.88 22.65 47.41 15.00
283 TRP N 32.71 19.57 49.93 15.00
283 TRP CA 31.49 19.17 50.65 15.00
283 TRP C 31.75 18.34 51.92 15.00
283 TRP 0 31.01 17.41 52.22 15.00
283 TRP CB 30.50 18.46 49.71 15.00
283 TRP CG 30.00 19.32 48.58 15.00
283 TRP CDl 30.06 20.69 48.50 15.00
283 TRP CD2 29.39 18.88 47.37 15.00
283 TRP NEl 29.52 21.11 47.32 15.00
283 TRP CE2 29.11 20.02 46.61 15.00
283 TRP CE3 29.06 17.62 46.87 15.00
283 TRP CZ2 28.51 19.94 45.35 15.00
283 TRP CZ3 28.47 17.54 45.62 15.00
283 TRP CH2 28.19 18.70 44.88 15.00

Claims

WHAT IS CLAIMED IS:
1. A method of inhibiting cathepsin K which comprises administering to a mammal in need thereof a compound that mimics the interaction of the propeptide with the residues of the active site of procathepsin K.
2. The method of claim 1 wherein the compound comprises:
(i) an oxygen atom which interacts with a hydrogen atom donated by the indole nitrogen of tryptophan 283 wherein the distance between these two atoms is 2.7-3.5A;
(ii) a hydrophobic group that interacts with tryptophan 283 wherein the distance between the centroid of said hydrophobic group and the centroid of the side chain atoms of tryptophan 283 is 4.10-7.10A; (iii) a hydrophobic group that interacts with tyrosine 166 , methionine
167, alanine 233, leucine 259, and leucine 308, creating a hydrophobic pocket, and has distance ranges between the centroid of said hydrophobic group and the centroids of the side chain atoms of the amino acid residues of said hydrophobic pocket which are tyrosine 166: 4.91- 5.91A, methionine 167: 5.74-6.74A, alanine 233: 4.15-5.15A, leucine 259: 6.18-7.18A, and leucine 308: 5.71-6.71 A; and
(iv) an amino group with a pKa of less than 7 or an oxygen atom, each of which interacts with a hydrogen atom donated by the amide nitrogen of glycine 264 wherein the distance between these two atoms is 2.7-3.5A.
3. The method of claim 2 wherein said hydrophobic group that interacts with said hydrophobic pocket is an isobutyl group.
4. The method of claim 1 wherein the compound is characterized by the interactions of the atoms of the procathepsin K active site and the atoms of the propeptide in the region of the acive site according to Table IV.
5. A composition comprising procathepsin K in crystalline form.
6. The composition according to claim 5 wherein the procathepsin K in crystalline form is characterized by structure coordinates according to Table I.
7. The composition according to claim 5 wherein procathepsin K has an active site cavity formed by the amino acids in Table IV.
8. The composition of claim 7 wherein said active site is characterized by the distance between the atoms of Table II and the bond angles between interresidue atoms of Table III.
9. A procathepsin K crystal.
10. The procathepsin K crystal according to claim 9 characterized by structure coordinates according to Table I.
11. An isolated, properly folded procathepsin K molecule or fragment thereof having a conformation comprising a catalytically active site formed by the residues listed in Table IV, said active site defined by the protein coordinates of Table I.
12. A peptide, peptidomimetic or synthetic molecule which binds with the active site cavity of procathepsin K according to claim 7.
13. A method of identifying an inhibitor compound capable of binding to, and inhibiting the proteolytic activity of, cathepsin K, said method comprising: introducing into a suitable computer program information defining an active site conformation of a procathepsin K molecule comprising a catalytically active site formed by the residues listed in Table IV, said active site defined by the protein coordinates of Table I, wherein said program displays the three-dimensional structure thereof; creating a three dimensional representation of the active site cavity in said computer program; displaying and superimposing the model of said test compound on the model of said active site; assessing whether said test compound model fits spatially into the active site; preparing said test compound that fits spatially into the active site; using said test compound in a biological assay for a protease characterized by said active site; and determining whether said test compound inhibits cathepsin K activity in said assay.
14. A peptide, peptidomimetic or synthetic molecule identified by the method of Claim 13.
15. A method of drug design comprising using the structural coordinates of a procathepsin K crystal to computationally evaluate a chemical entity for associating with the active site of cathepsin K.
16. The method according to claim 15, wherein said entity is a competitive or non-competitive inhibitor of cathepsin K.
17. A method for identifying inhibitors which competitively bind to the active site of a cathepsin K molecule or fragment thereof said method comprising the steps of: providing the coordinates of the active site of procathepsin K characterized by the residues listed in Table IV to a computerized modeling system; identifying compounds which will bind to the structure; and screening the compounds identified for protease inhibitory bioactivity.
18. A method of identifying inhibitors of cathepsin K said method comprising the steps of: providing the coordinates of the atoms of the procathepsin K active site and the coordinates of the atoms of the propeptide in the region of the active site according to Table IV to a computerized modeling system, wherein said system displays the three-dimensional structure thereof; displaying and superimposing a model of a test compound on the model of the propeptide; assessing whether said test compound interacts with the active site of procathepsin K in a manner similar to that of the propeptide; preparing said test compound that interacts with the active site of procathepsin K in a manner similar to that of the propeptide; and testing said test compound for cathepsin K inhibitory activity.
PCT/US1998/014634 1998-08-14 1998-08-14 Method of inhibiting cathepsin k Ceased WO2000009653A2 (en)

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WO2002086115A1 (en) 2001-04-20 2002-10-31 The University Of Georgia Research Foundation, Inc. Active site inactivators
US7071184B2 (en) 2000-03-21 2006-07-04 Smithkline Beecham Corporation Protease inhibitors
WO2007012180A1 (en) * 2005-07-26 2007-02-01 Merck Frosst Canada Ltd. Papain family cysteine protease inhibitors for the treatment of parasitic diseases
US7405209B2 (en) 1998-12-23 2008-07-29 Smithkline Beecham Corporation Protease inhibitors
US8642799B2 (en) 2007-11-29 2014-02-04 Merck Canada Inc. Cysteine protease inhibitors for the treatment of parasitic diseases
EP2719700A1 (en) 2008-01-09 2014-04-16 Amura Therapeutics Limited Tetrahydrofuro(3,2-b)pyrrol-3-one derivatives as inhibitors of cysteine proteinases

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SK88997A3 (en) * 1995-10-30 1998-05-06 Smithkline Beecham Corp Method of inhibiting cathepsin k
SK56798A3 (en) * 1995-10-30 1998-12-02 Smithkline Beecham Corp Protease inhibitors, pharmaceutical composition containing them and their use

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7405209B2 (en) 1998-12-23 2008-07-29 Smithkline Beecham Corporation Protease inhibitors
US7071184B2 (en) 2000-03-21 2006-07-04 Smithkline Beecham Corporation Protease inhibitors
US7563784B2 (en) 2000-03-21 2009-07-21 Smithkline Beecham Corporation Protease inhibitors
WO2002086115A1 (en) 2001-04-20 2002-10-31 The University Of Georgia Research Foundation, Inc. Active site inactivators
EP1379634A4 (en) * 2001-04-20 2004-09-08 Univ Georgia Res Found Active site inactivators
WO2007012180A1 (en) * 2005-07-26 2007-02-01 Merck Frosst Canada Ltd. Papain family cysteine protease inhibitors for the treatment of parasitic diseases
US8642799B2 (en) 2007-11-29 2014-02-04 Merck Canada Inc. Cysteine protease inhibitors for the treatment of parasitic diseases
EP2719700A1 (en) 2008-01-09 2014-04-16 Amura Therapeutics Limited Tetrahydrofuro(3,2-b)pyrrol-3-one derivatives as inhibitors of cysteine proteinases

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JP2003527819A (en) 2003-09-24
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