Patent Document

CROSS-REFERENCE TO RELATED APPLICATIONS  
       [0001]     This application is a divisional of U.S. patent application Ser. No. 10/444,257; filed May 21, 2003 which claims the benefit of U.S. provisional patent application No. 60/383,391 filed May 24, 2002, the contents of each of which are hereby incorporated by reference in their entireties. 
     
    
     BACKGROUND OF THE INVENTION  
       [0002]     1. Field of Invention  
         [0003]     The present invention pertains to a modified tumor necrosis factor-alpha converting enzyme (TACE). The present invention further pertains to generating a crystal of the modified TACE protein in protein-ligand complex with a selected inhibitor for use in structure based rational drug design. In addition, the present invention pertains to methods of using these proteins and crystals to identify compounds that can modulate the enzymatic activity of TACE.  
         [0004]     2. Background  
         [0005]     Tumor necrosis factor-alpha (TNF-alpha) plays a major role in the immune and inflammatory responses in mammals [Qi and Pekala,  Proc Soc Exp Biol Med.  223(2): 128-135 (2000)]. First isolated from the serum of rabbits that had been treated with endotoxin, TNF-alpha was named for its ability to trigger the hemorrhagic necrosis of specific transplantable tumors [Old,  Science  230:630-633 (1985)]. Subsequently, TNF-alpha was found to be identical to cachetin, a protein that is intimately involved in cachexia, the wasting disease prevalent in AIDS and cancer [Beutler et al.,  J. Exp. Med.,  161:984-995 (1985)].  
         [0006]     TNF-alpha can bind to two distinct cell membrane receptors (TNFR1 and TNFR2) to transduce intercellular signals to a variety of different target cells in a number of different tissues [Qi and Pekala,  Proc Soc Exp Biol Med.  223(2): 128-135 (2000)]. Though a central participant in several key cellular processes, TNF-alpha also functions deleteriously as a mediator of insulin resistance in diabetes mellitus [Hotamisligil and Spiegelman,  Diabetes  43:1271-1278 (1994)]. In addition, an over-abundance of the active form of TNF-alpha has been linked to adverse symptoms in a number of disease states, including in rheumatoid arthritis, Crohn&#39;s disease, sepsis, and cachexia. Since there are presently no effective treatments for these conditions, there remains a need to find new drugs that can be used to modulate the effective physiological concentration of the active form of TNF-alpha.  
         [0007]     TNF-alpha is transcribed as a transmembrane protein having a monomeric molecular weight of 26 kilodaltons [Shirai et al.,  Nature  313:803-806 (1985)]. A metalloprotease, tumor necrosis factor-alpha converting enzyme (TACE) cleaves the membrane-associated form of TNF-alpha at a specific site of the protein, converting it to its corresponding soluble form [Black et al.,  Nature  385:729-733 (1997); Moss et al.,  Nature  385:733-736 (1997); U.S. Pat. No. 5,830,742, Issued Nov. 3, 1998; U.S. Pat. No. 6,013,466, Issued Jan. 11, 2000, the contents of which are hereby incorporated by reference in their entireties]. The soluble form of TNF-alpha then associates as a homotrimer of three 17 kilodalton monomers [Kriegler et al.,  Cell,  53:45-53 (1988)]. Although the membrane-associated form of TNF-alpha appears to be active, it is the proteolyzed soluble form that is responsible for the mortality associated with endotoxic shock [Gearing et al.,  Nature  370:555-558 (1994)]. Thus, reducing the circulating concentration of active TNF-alpha appears to be critical to alleviate the harmful side effects caused by this cytokine. One means for achieving this reduction in concentration of soluble TNF-alpha is to inhibit the TACE protease.  
         [0008]     TACE, also referred to as ADAM 17 and CD156q, is a zinc endopeptidase that is a member of the “A Disintegrin And Metalloprotease” (ADAM) family of metalloproteases [Schlondorff and Blobel,  J. Cell Sci.,  112:3603-3617 (1999); Black,  Intern. J. Biochem. Cell Biol  34:1-5 (2002); U.S. Pat. No. 5,830,742, Issued Nov. 3, 1998, the contents of all of which are hereby incorporated by reference in their entireties]. A type I transmembrane protein, TACE comprises (i) an extracellular region having an N-terminal signal peptide, (ii) a pro domain, (iii) a zinc-dependent catalytic domain, (iv) a disintegrin domain, (v) an EGF-like domain, (vi) a crambin-like domain, (vii) a transmembrane helix and (viii) an intracellular C-terminal tail [see WO9940182, Published Aug. 12, 1999]. More recently, the EGF-like and the crambin-like domains have been grouped together and re-named as a cysteine-rich domain [Black,  Intern. J. Biochem. Cell Biol  34:1-5 (2002)].  
         [0009]     Since TACE is a protease, the portion of the enzyme that is critical for drug discovery is its catalytic domain. The catalytic domain (TCD) of TACE comprises 263 amino acid residues preceded by a furin cleavage site (residues 211-214). The pro domain comprises a cysteine that interacts with the zinc molecule at the active-site preventing proteolytic action. Therefore, this cysteine must be displaced in order to generate an active protease [Black, Intern.  J. Biochem. Cell Biol  34:1-5 (2002)].  
         [0010]     Zask et al. have prepared a compilation of inhibitors of metalloproteinases [ Curr. Pharm. Des.,  2:624-661 (1996), the contents of which are hereby incorporated by reference in their entireties], and more recently, Letavic et al. has disclosed several specific inhibitors of TACE [ Biorgan. &amp; Medic. Chem Lett.  12:1387-1390 (2002), the contents of which are hereby incorporated by reference in their entireties]. To date, however, none of these has proven to be useful in the treatment of conditions related to an over-abundance of soluble TNF-alpha.  
         [0011]     Three-dimensional structures of two different TACE-inhibitor complexes have been obtained via X-ray crystallographic analyses [Letavic et al.,  Biorgan. &amp; Medic. Chem Lett.  12:1387-1390 (2002); WO9940182, Published Aug. 12, 1999, U.S. application Ser. No. 09/117,476, filed Jan. 27, 1999, the contents of which are all hereby incorporated by reference in their entireties]. Importantly, however, the conditions for preparing the two TACE-inhibitor complexes were significantly different. These results suggest that new crystallization conditions may be required for every different TACE-ligand complex. Determining crystallization conditions, de novo can be extremely time-consuming. Moreover, such a requirement severely hampers progress in identifying new and more potent inhibitors of TACE which are necessary for developing safe and effective drugs to ameliorate the deleterious effects due to an overabundance of the soluble form of TNF-alpha.  
         [0012]     Therefore, there is need to provide methods for performing X-ray crystallographic structural determinations on multiple TACE protein-ligand complexes without having to determine crystallization conditions, de novo. In addition, there is a need to obtain X-ray diffractable crystals of the TACE catalytic domain that are stable. Furthermore, there is a need to obtain crystals of the TACE catalytic domain that are amenable to ligand exchange.  
         [0013]     The citation of any reference herein should not be construed as an admission that such reference is available as “prior art” to the instant application.  
       SUMMARY OF THE INVENTION  
       [0014]     The present invention discloses that the native TACE protein unexpectedly undergoes autoproteolysis at the high protein concentrations required for X-ray crystallography. The present invention further identifies the peptide bond between amino acid residues Tyr352 and Val353 (Y 352 -V 353 ) of SEQ ID NO: 2 as the specific cleavage site.  
         [0015]     Therefore, the present invention provides a polypeptide comprising a modified TACE catalytic domain that is significantly less susceptible to autoproteolysis than the native TACE. The modified TACE of the present invention imparts improved stability to the protease under the conditions employed to generate TACE crystals. The present invention also uniquely enables X-ray crystallographic structural determinations to be performed on multiple TACE protein-ligand complexes in rapid succession. This ability to rapidly generate three-dimensional structures of TACE protein-ligand complexes is critical for a successful structure based rational drug design program.  
         [0016]     Indeed, the structural information generated using the compositions and methods of the present invention greatly facilitates the identification of new and more potent inhibitors of the TACE protease. Selected inhibitors, in turn, become lead candidates in the development of drugs that will be useful for counteracting the harmful effects due to an overabundance of the soluble form of TNF-alpha, i.e., drugs that can be used in the treatment of rheumatoid arthritis, Crohn&#39;s disease, sepsis, and/or cachexia.  
         [0017]     One aspect of the present invention provides a polypeptide comprising the amino acid sequence of SEQ ID NO: 8. In one such embodiment, the polypeptide consists essentially of the amino acid sequence of SEQ ID NO: 8. In a particular embodiment, the amino acid residue at position 139 of SEQ ID NO: 8 is serine. 1  In another embodiment of this type, the amino acid residue at position 139 of SEQ ID NO: 8 is alanine. In a preferred embodiment, the amino acid residue at position 139 of SEQ ID NO: 8 is glycine (denoted as “vgTACE” in the Example below, and has the amino acid sequence of SEQ ID NO: 20).    1 The amino acid sequences of the TACE catalytic domain and the modified TACE catalytic domain are SEQ ID NOs: 6 and 8, respectively. Position 139 of SEQ ID NOs: 6 and 8 is equivalent to position 353 of SEQ ID NO: 2, the amino acid sequence of the TACE polypeptide comprising the Pre, Pro and catalytic domains.    
         [0018]     In a related embodiment, the present invention provides a polypeptide comprising a modified TACE catalytic domain comprising at least 95% identity with the amino acid sequence of SEQ ID NO: 8. Preferably, this polypeptide catalyzes the proteolytic cleavage of SEQ ID NO: 17 and/or binds to N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide and/or binds to N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH. In one such embodiment, the polypeptide consists essentially of a modified TACE catalytic domain having at least 95% identity with the amino acid sequence of SEQ ID NO: 8. In one specific embodiment of this type, the amino acid residue at position 139 of SEQ ID NO: 8 is alanine. In another embodiment, the amino acid residue at position 139 of SEQ ID NO: 8 is glycine. In yet another embodiment, the amino acid residue at position 139 of SEQ ID NO: 8 is serine.  
         [0019]     The present invention also provides the full-length TACE polypeptide and fragments thereof that comprise the TACE modified catalytic domain. In one such embodiment the TACE protein comprises (i) the pre domain, (ii) the pro domain, and (iii) the modified catalytic domain. In another embodiment, the fragment of the TACE polypeptide comprises the pro domain, and the modified catalytic domain.  
         [0020]     Chimeric proteins comprising the polypeptides of the present invention are also part of the present invention. In a particular embodiment, the chimeric TACE is a fusion protein comprising (i) the pre domain, (ii) the pro domain, (iii) the modified catalytic domain and (iv) a polyhistidine tag. In another embodiment, the chimeric protein comprises the modified catalytic domain and a polyhistidine tag. In a preferred embodiment, the polyhistidine tag further comprises a glycyl-seryl (i.e., Gly-Ser) linker.  
         [0021]     The present invention further provides nucleic acids that encode the polypeptides and chimeric proteins of the present invention (see e.g., Table 1 below). In a particular embodiment, the nucleic acid encodes a polypeptide having the amino acid of SEQ ID NO: 8. The present invention further provides expression vectors that comprise the nucleic acids of the present invention and a transcriptional control sequence. Preferably the nucleic acids of the present invention are operatively linked to the transcriptional control sequences in the expression vectors. Host cells comprising the expression vectors are also part of the present invention.  
         [0022]     In addition, the present invention provides methods for producing the above-mentioned polypeptides. One such embodiment comprises culturing a host cell of the present invention that produces the polypeptides. Methods for purifying the resulting recombinant polypeptides are also included in the present invention, as are the purified recombinant polypeptides.  
         [0023]     Crystals, each comprising one of the protein-ligand complexes of the present invention, are also contemplated. Preferably, such crystals effectively diffract X-rays for the determination of the atomic coordinates of the protein-ligand complex to a resolution of greater than 5.0 Angstroms. More preferably, a crystal of the present invention effectively diffracts X-rays for the determination of the atomic coordinates to a resolution of greater than 3.5 Angstroms. Even more preferably, a crystal of the present invention effectively diffracts X-rays for the determination of the atomic coordinates equal to or greater than 3.0 Angstroms. Still more preferably, a crystal of the present invention effectively diffracts X-rays for the determination of the atomic coordinates equal to or greater than 2.5 Angstroms, and, yet even more preferably, equal to or greater than 2.0 Angstroms. Most preferably, a crystal of the present invention effectively diffracts X-rays for the determination of the atomic coordinates equal to or greater than 1.5-1.7 Angstroms.  
         [0024]     In a particular embodiment, the crystal comprises a protein-ligand binding complex in which the polypeptide comprises the amino acid sequence of SEQ ID NO: 8. In another embodiment, the crystal comprises a polypeptide comprising a protein-ligand binding complex in which the polypeptide comprises a modified TACE catalytic domain having at least 95% identity with the amino acid sequence of SEQ ID NO: 8. Preferably this polypeptide catalyzes the proteolytic cleavage of SEQ ID NO: 17, and/or binds to the compound, N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide and/or binds to N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH. In one particular embodiment of this type, the crystal of the present invention has the space group P2 1 2 1 2 1 , with unit cell dimensions of: a=73, b=75, c=103 Angstroms.  
         [0025]     Another aspect of the present invention provides methods for obtaining a crystal comprising a protein-ligand complex between a substitute ligand and a modified TACE catalytic domain. One such method comprises incubating (e.g., soaking) an excess of a substitute ligand with a crystal comprising a modified TACE catalytic domain and an initial ligand in a protein-ligand binding complex. The incubation is performed under the appropriate conditions and for a sufficient time period for the substitute ligand to replace the initial ligand in the protein-ligand complex. A crystal comprising the protein-ligand complex between the substitute ligand and the modified TACE catalytic domain is thus, obtained. The modified TACE catalytic domain can be part of a larger polypeptide (e.g., the full-length TACE or a chimeric protein). In a preferred embodiment, the modified TACE catalytic domain comprises the amino acid sequence of SEQ ID NO: 8. In a one such embodiment, the modified TACE catalytic domain comprises the amino acid sequence of SEQ ID NO: 20. In a preferred embodiment, the initial ligand is N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide. In a particular embodiment of this type, the substitute ligand is N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH. The present invention further provides the crystals that comprise protein-ligand binding complexes that have had their initial ligand replaced with a substitute ligand.  
         [0026]     In an alternative embodiment, the present invention provides a method for identifying an agent for use as an inhibitor of TACE. One such embodiment comprises obtaining a set of atomic coordinates that define the three-dimensional structure of the protein-ligand binding complex from a crystal of the present invention. A potential agent is then selected by performing rational drug design with the atomic coordinates obtained. Preferably, the selection is performed in conjunction with computer modeling. The potential agent is contacted with a proteolytic polypeptide that comprises the catalytic domain of TACE, or alternatively, an active fragment thereof. The catalytic activity of the proteolytic polypeptide is then determined in a TACE activity assay. A potential agent is identified as an agent that inhibits TACE when there is a decrease in the activity of the proteolytic polypeptide in the presence of the agent relative to in its absence.  
         [0027]     In a related embodiment, a method of identifying a compound that is predicted to inhibit TACE is provided. One such embodiment comprises using the atomic coordinates in Table 3 to define the structure of the complex between the modified catalytic domain of TACE and N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH and/or to define the structure of a portion of that complex. The portion of the complex defined is required to comprise sufficient structural information to enable the identification of a given compound as a potential inhibitor. A compound then is identified as one predicted to inhibit TACE through the use of the defined structure. A particular embodiment of this type further comprises contacting the compound with a proteolytic polypeptide comprising the catalytic domain of TACE or an active fragment thereof and determining the catalytic activity of the proteolytic polypeptide in a TACE activity assay. A potential agent is identified as an agent that inhibits TACE when there is a decrease in the activity of the proteolytic polypeptide in the presence of the agent relative to in its absence.  
         [0028]     The present invention further provides a computer comprising a three-dimensional representation of a protein-ligand complex between a modified TACE catalytic domain and N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH in computer memory. One such embodiment comprises: (i) a machine-readable data storage medium comprising a data storage material encoded with machine-readable data comprising the atomic coordinates of Table 3; (ii) a working memory for storing instructions for processing the machine-readable data; and (iii) a central-processing unit coupled to the working memory and to the machine-readable data storage medium for processing the machine readable data into a three-dimensional representation. Preferably a display is also provided that is coupled to the central-processing unit for visualizing the three-dimensional representation of the protein-ligand complex.  
         [0029]     Accordingly, it is a principal object of the present invention to provide an active TACE catalytic domain that can form a stable X-ray diffractable crystal.  
         [0030]     It is a further object of the present invention to provide a way for exchanging ligands of a TACE catalytic domain in a crystal.  
         [0031]     It is a further object of the present invention to provide multiple crystals of the TACE catalytic domain each comprising a different protein-ligand complex.  
         [0032]     It is a further object of the present invention to provide an effective way of performing structure based rational drug design with TACE.  
         [0033]     It is a further object of the present invention to provide drugs to treat conditions due to an overabundance of the soluble form of TNF-alpha.  
         [0034]     These and other aspects of the present invention will be better appreciated by reference to the following drawing and Detailed Description. 
     
    
     BRIEF DESCRIPTION OF THE DRAWING  
       [0035]      FIG. 1  depicts a schematic of a computer comprising a central processing unit (CPU), a working memory, a mass storage memory, a display terminal, and a keyboard that are interconnected by a conventional bidirectional system bus. The computer can be used to display and manipulate the structural data of the present invention. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0036]     Although entirely independent of any particular mechanism, the present invention was conceived following the unexpected discovery by the present inventors that the native TACE protein undergoes autoproteolysis at the high protein concentrations required for X-ray crystallographic analysis. The subsequent identification by the inventors of an autoproteolysis site between Y 352 -V 353  of SEQ ID NO: 2, raised the possibility that the replacement of either one or both of these amino acid residues might lead to a TACE protein that was resistant to autoproteolysis. Insertion(s) or deletion(s) of amino acid residues adjacent to the Y 352 -V 353  cleavage site might also lead to a TACE polypeptide that is resistant to autoproteolyisis. However, inserting or deleting amino acid residues adjacent to the Y 352 -V 353  cleavage site could create conformational changes in the protein that significantly reduce its enzyme activity and/or stability. Similarly, since Tyr352 of SEQ ID NO: 2 is located within a hydrophobic pocket that is close to the active site of the TACE protease, modification of this amino acid residue also might effect the enzymatic activity and/or protein stability. In direct contrast, Val353 of SEQ ID NO: 2 is located on the enzyme surface with its side chain exposed to the solvent, and so modification of the valine side chain might be less traumatic to the enzyme structure. Therefore, modification of Val353 is preferred over either modifying Tyr352, or inserting or deleting amino acid residues adjacent to the Y 352 -V 353  cleavage site. These latter two alternatives, however, remain part of the present invention.  
         [0037]     Indeed, as disclosed herein, substitution of the hydrophobic valine side chain with either serine or glycine significantly reduces autoproteolysis, and dramatically improved the stability of the protein, without significantly altering the proteolytic activity of the TACE enzyme. In a preferred embodiment, the modified TACE catalytic domain contains an amino acid change at amino acid residue 353 of SEQ ID NO: 2. In one such embodiment, Val353 is replaced with a glycyl residue. In another embodiment, Val353 is replaced with a seryl residue. Substitutions of the nonpolar side chain of valine with alternative non-hydrophobic side chains can also prevent auto-proteolysis, and such substitutions are also part of the present invention. In addition, in order to remove N-glycosylation sites it is preferable as exemplified below, that Ser266 be replaced e.g., with an alanyl residue, and Asp452 be replaced e.g., with a glutaminyl residue.  
         [0038]     The present invention further provides crystals comprising a protein-ligand complex of a polypeptide that comprises a modified TACE catalytic domain. The three-dimensional structure of a protein-ligand complex comprising a modified TACE catalytic domain bound to N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH is provided in the Example below (see Table 3 which lists the atomic coordinates).  
         [0039]     N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH which is commercially available, e.g., from Chem-Impex International, Wood Dale II, (product code 09538), has the following chemical structure:  
                         
 
         [0040]     Structure based rational drug design is the most efficient method of drug development. In one common paradigm, a three dimensional structure of a protein-ligand complex is determined, and potential antagonists (e.g., inhibitors) of the protein (e.g., potential drugs) are identified and/or designed with the aid of computer modeling [Bugg et al.,  Scientific American , Dec.: 92-98 (1993); West et al.,  TIPS,  16:67-74 (1995); Dunbrack et al.,  Folding &amp; Design,  2:27-42 (1997)]. The drug candidates are selected and assayed. The most promising drug candidates are identified, and then incubated in excess with crystals of a protein-ligand complex to replace the initial ligand. The three-dimensional structure of the new protein-ligand complex is then determined, and new potential antagonists. of the protein are identified and/or designed with the aid of computer modeling. This process can then be continued in successive iterations until a lead drug candidate is identified.  
         [0041]     Heretofore, the ability to perform structure based rational drug design with TACE was severely hampered because only two TACE protein-ligands complexes were known to form an X-ray quality crystal, [Maskos et. al.,  Proc. Natl. Acad. Sci. USA  95:3408-3412 (1998); Letavic et al.,  Biorgan. &amp; Medic. Chem Lett.  12:1387-1390 (2002)], and these crystals were not reported to be capable of ligand exchange. As disclosed herein, the present invention overcomes this problem by providing crystals of the modified TACE catalytic domain that are conducive to ligand exchange.  
         [0042]     As used herein the following terms shall have the definitions set out below:  
         [0043]     As used herein the term “polypeptide” is used interchangeably with the term “protein” and is further meant to encompass peptides. Therefore, as used herein, a polypeptide is a polymer of two or more amino acids joined together by peptide linkages. Preferably, a polypeptide is a polymer comprising twenty or more amino acid residues joined together by peptide linkages.  
         [0044]     As used herein a polypeptide “consisting essentially of” or that “consists essentially of” a specified amino acid sequence is a polypeptide that (i) retains the general characteristics of a polypeptide comprising that amino acid sequence, e.g., the activity of the polypeptide, and (ii) further comprises the identical amino acid sequence, except it consists of plus or minus 10% (or a lower percentage), preferably plus or minus 5% (or a lower percentage), and more preferably plus or minus 2.5% (or a lower percentage) of the amino acid residues.  
         [0045]     As used herein a “modified TACE catalytic domain” is a TACE catalytic domain that has been modified to resist and/or prevent autocatalysis. Preferably, at least one of the two critical amino acid residues at the autoproteolytic site of TACE, i.e., Tyr352 and Val353 of SEQ ID NO: 2, has been replaced. More preferably, a modified TACE catalytic domain has the Val353 residue replaced with a non-hydrophobic amino acid residue.  
         [0046]     As used herein a “non-hydrophobic amino acid” is an amino acid that is not hydrophobic. The genus of non-hydrophobic amino acids specifically does not include leucine, isoleucine, valine, methionine, tryptophan, and phenylalanine.  
         [0047]     As used herein a “polypeptide comprising a modified TACE catalytic domain”, can be (i) the full length TACE protein, (ii) a fragment of the TACE protein that includes the modified TACE catalytic domain e.g., the pro and catalytic domain, (iii) the modified TACE catalytic domain alone, or (iv) a chimeric protein which comprises any of the above.  
         [0048]     As used herein a “proteolytic polypeptide” of the present invention is a polypeptide that is capable of catalyzing the proteolytic cleavage of a substrate of the native TACE protease. A proteolytic polypeptide of the present invention minimally comprises an active fragment of the TACE catalytic domain that retains proteolytic activity. A proteolytic polypeptide of the present invention can be a chimeric protein.  
         [0049]     As used herein an “active fragment” of the catalytic domain of TACE is a fragment of the catalytic domain of TACE and/or modified TACE catalytic domain that retains at least about 10%, preferably at least about 20%, and more preferably at least about 25% of the proteolytic activity of the full-length TACE protease. Preferably, the active fragment retains at least about 25%, more preferably at least about 50%, and even more preferably at least about 75% of the amino acid residues of the catalytic domain of TACE having the amino acid sequence of SEQ ID NO: 6. More preferably, the amino acid sequence of the active fragment of the TACE catalytic domain has at least about 95% identity to the corresponding amino acid residues of SEQ ID NO: 6.  
         [0050]     As used herein the term “chimeric” protein is meant to include “fusion proteins”. “Chimeric” proteins of the present invention comprise at least a portion of a non-TACE protein joined via a peptide bond to at least a portion of a TACE polypeptide. Chimeric proteins can have additional structural, regulatory, or catalytic properties. In a particular embodiment the chimeric protein functions as a means of detecting and/or isolating the TACE polypeptide or fragment thereof after the recombinant nucleic acid is expressed. Non-TACE amino acid sequences are preferably either amino- or carboxy-terminal to the TACE sequence.  
         [0051]     As used herein one amino acid sequence is 100% “identical” to a second amino acid sequence when the amino acid residues of both sequences are identical. Accordingly, an amino acid sequence is 50% “identical” to a second amino acid sequence when 50% of the amino acid residues of the two amino acid sequences are identical. The sequence comparison is performed over a contiguous block of amino acid residues comprised by the TACE polypeptide or the portion of the TACE polypeptide being compared, e.g., the modified catalytic domain (SEQ ID NO: 8). In a preferred embodiment, selected deletions or insertions that could otherwise alter the correspondence between the two amino acid sequences are taken into account.  
         [0052]     As used herein, DNA and protein sequence percent identity can be determined using C, MacVector 6.0.1, Oxford Molecular Group PLC (1996) and the Clustal W algorithm with the alignment default parameters, and default parameters for identity. These commercially available programs can also be used to determine sequence similarity using the same or analogous default parameters. Alternatively, an Advanced Blast search under the default filter conditions can be used, e.g., using the GCG (Genetics Computer Group, Program Manual for the GCG Package, Version 7, Madison, Wis.) pileup program using the default parameters.  
         [0053]     As used herein a “nucleic acid” refers to the phosphate ester polymeric form of ribonucleosides (adenosine, guanosine, uridine or cytidine; “RNA molecules”) or deoxyribonucleosides (deoxyadenosine, deoxyguanosine, deoxythymidine, or deoxycytidine; “DNA molecules”), or any phosphoester analogs thereof, such as phosphorothioates and thioesters, in either single stranded form, or a double-stranded helix. Double stranded DNA-DNA, DNA-RNA and RNA-RNA helices are possible. When referring to a nucleic acid that is double stranded both the “sense” strand and the complementary “antisense” strand are intended to be included. Thus a nucleic acid that is hybridizable to SEQ ID NO: 1, for example, can be either hybridizable to the “sense” strand of SEQ ID NO: 1, which is particularly listed in the SEQUENCE LISTING, or to the “antisense” strand which can be readily determined from that SEQUENCE LISTING.  
         [0054]     A DNA “coding sequence” is a double-stranded DNA sequence that is transcribed and translated into a polypeptide in a cell in vitro or in vivo when placed under the control of appropriate regulatory sequences. The boundaries of the coding sequence are determined by a start codon at the 5′ (amino) terminus and a translation stop codon at the 3′ (carboxyl) terminus. A coding sequence can include, but is not limited to, prokaryotic sequences, cDNA from eukaryotic MRNA, genomic DNA sequences from eukaryotic (e.g., mammalian) DNA, and even synthetic DNA sequences. If the coding sequence is intended for expression in a eukaryotic cell, a polyadenylation signal and transcription termination sequence will usually be located 3′ to the coding sequence.  
         [0055]     Transcriptional and translational control sequences are DNA regulatory sequences, such as promoters, enhancers, terminators, and the like, that provide for the expression of a coding sequence in a host cell. In eukaryotic cells, polyadenylation signals are control sequences.  
         [0056]     A “promoter sequence” is a DNA regulatory region capable of binding RNA polymerase in a cell and initiating transcription of a downstream (3′ direction) coding sequence. For purposes of defining the present invention, the promoter sequence is bounded at its 3′ terminus by the transcription initiation site and extends upstream (5′ direction) to include the minimum number of bases or elements necessary to initiate transcription at levels detectable above background. Within the promoter sequence will be found a transcription initiation site (conveniently defined for example, by mapping with nuclease S1), as well as protein binding domains (consensus sequences) responsible for the binding of RNA polymerase.  
         [0057]     A coding sequence is “under the control” of transcriptional and translational control sequences in a cell when RNA polymerase transcribes the coding sequence into mRNA, which can then be trans-RNA spliced and translated into the protein encoded by the coding sequence.  
         [0058]     A nucleic acid sequence is “operatively linked” to an expression control sequence when the expression control sequence controls or regulates the transcription and translation of that nucleic acid sequence. The term operatively linked includes having an appropriate start signal.  
         [0059]     A “heterologous nucleotide sequence” as used herein is a nucleotide sequence that is added to a nucleotide sequence of the present invention by recombinant methods to form a nucleic acid that is not naturally formed in nature. Such nucleic acids can encode chimeric proteins. Alternatively, a heterologous nucleotide sequence can contain a nucleic acid regulatory sequence. Thus a heterologous nucleotide sequence can comprise non-coding sequences including restriction sites, regulatory sites, promoters and the like. In still another embodiment the heterologous nucleotide can function as a means of detecting a nucleotide sequence of the present invention. The present invention provides heterologous nucleotide sequences that when combined with nucleotide sequences encoding the TACE proteins, and fragments thereof, are necessary and sufficient to encode all of the chimeric proteins of the present invention.  
         [0060]     As used herein the phrases “structure based rational drug design”, “structure based drug design”, “structure assisted drug design” and “rational drug design” are used interchangeably. These phrases are meant to convey a particular method of identifying and/or designing a ligand (preferably an inhibitor) for a specific target protein that includes the use of the three-dimensional structure of that protein and/or its corresponding protein-ligand complex.  
         [0061]     The phrase “binding to” in regard to a ligand binding to a polypeptide is used herein to include any or all such specific interactions that lead to a protein-ligand binding complex. This can include processes such as covalent, ionic, hydrophobic and hydrogen bonding, but does not include non-specific associations such solvent preferences.  
         [0062]     As used herein a “ligand” of a polypeptide is a compound that binds to the polypeptide in a protein-ligand binding complex. In a specific embodiment of the present invention the polypeptide has an enzymatic activity and the ligand inhibits that activity when bound to the polypeptide in a protein-ligand binding complex. Such a ligand is also termed an “inhibitor”.  
         [0063]     As used herein the term “initial ligand” denotes a ligand in a protein-ligand complex that is, or can be displaced by a “substitute ligand”.  
         [0064]     As used herein, a “protein-ligand binding complex” is a specific association between a polypeptide and the compound that binds to it. In a preferred embodiment of the present invention, the ligand is an inhibitor of the polypeptide. In a particular embodiment of this type, the binding of the inhibitor to the polypeptide occurs at the active site of the polypeptide.  
         [0065]     As used herein “incubating a ligand with a crystal” is used interchangeably with “soaking a crystal with a ligand”. Incubating a ligand with a crystal is the contacting of a ligand with a crystal of a polypeptide under the appropriate conditions and for a sufficient time period (e.g., hours to several days) for the ligand to bind to the crystalline polypeptide and form a crystalline protein-ligand complex. Such incubating can further and/or alternatively, include contacting an excess of a substitute ligand with a crystal of a protein-ligand complex under the appropriate conditions and for a sufficient time period (e.g., hours to several days) for the substitute ligand to replace the initial ligand and form the new crystalline protein-ligand complex.  
         [0066]     As used herein the terms “displacing”, “replacing”, and “exchanging” are used interchangeably in regard to the substitution of one ligand in a protein-ligand complex for another.  
         [0067]     As used herein an “excess of a substitute ligand” is an amount of that ligand that is sufficient to replace 80% or more, and preferably 90% or more, of the initial ligand in a protein-ligand complex. In a particular embodiment of this type, the concentration of the substitute ligand is about ten-fold higher than the concentration of the protein-ligand complex. In a preferred embodiment, the concentration of the substitute ligand is about one hundred-fold higher than the concentration of the protein-ligand complex.  
         [0068]     As used herein the term “X-ray diffractable crystal” is a crystal of a compound that yields a discernable diffraction pattern when subjected to 0.5 to 2.5 Å incident X-ray radiation.  
         [0069]     As used herein an “X-ray quality crystal” is an X-ray diffractable crystal that can yield meaningful structural data of its crystalline composition when subjected to X-ray crystallographic analysis.  
         [0070]     As used herein, and unless otherwise specified, the terms “agent”, “potential drug”, “compound”, or “test compound” are used interchangeably, and refer to chemicals that have or potentially have a use as an inhibitor of the proteolytic activity of TACE. Preferably such agents include drugs for the treatment or prevention of a disease and/or condition involving the proteolytic action of TACE. Therefore, such agents may be used, as described herein, in drug assays and drug screens and the like.  
         [0071]     As used herein a “small organic molecule” is an organic compound [or organic compound complexed with an inorganic compound (e.g., metal)] that has a molecular weight of less than 3 Kd.  
         [0072]     As used herein the terms “approximately” and “about” are used to signify that a value is within twenty percent of the indicated value i.e., an amino acid sequence containing “approximately” 260 amino acid residues can contain between 208 and 312 amino acid residues.  
       Nucleic Acids Encoding TACE  
       [0073]     Obtaining and/or constructing a cDNA that encodes a polypeptide comprising a modified TACE facilitates the production of the large quantities of protein required to perform X-ray crystallographic analysis. Since the sequence of the native protein is known [see U.S. Pat. No. 6,013,466, Issued Jan. 11, 2000, the contents of which are hereby incorporated by reference in their entireties], a cDNA encoding the modified protease can be readily obtained.  
         [0074]     To express a recombinant protein of the present invention in a host cell, an expression vector can be constructed comprising the corresponding cDNA. The present invention therefore, provides expression vectors containing nucleic acids encoding polypeptides comprising the modified TACE catalytic domains of the present invention. Due to the degeneracy of nucleotide coding sequences, other DNA sequences which encode substantially the same amino acid sequence as a nucleic acid encoding a polypeptide comprising the modified TACE catalytic domain of the present invention may be used in the practice of the present invention. These include, but are not limited to, allelic genes, homologous genes from other species, which are altered by the substitution of different codons that encode the same amino acid residue within the sequence, thus producing a silent change. Host cells comprising the expression vectors of the present invention are also provided.  
         [0075]     Cloning of cDNAs and expression of their corresponding recombinant proteins have become a routine laboratory exercise [see Sambrook and Russell,  Molecular Cloning, A laboratory Manual,  3 rd    edition , Cold Spring Harbor Laboratory Press, Cold Spring Harbor L.I. (2000), the contents of which are hereby incorporated by reference in their entireties]. The use of a Baculovirus recombination system and Sf9 host cells is exemplified below. Purification of recombinant proteins has also become a routine laboratory exercise. In the present case, the modified TACE protein was cloned and expressed as the pro-protein. The pre and pro domains were cleaved during protein expression and secretion by the cells. The catalytic domain was then purified (see Example below).  
         [0076]     The nucleotide sequence for open reading frame of TACE with a GS linker, a polyHis tag (H6), and stop codon is shown below. The nucleic acid sequence encoding: (i) the pre domain is underlined, (ii) the pro domain is under-dashed, (iii) the catalytic domain is unmarked and (iv) the GS linker and the polyHis tag (H6) are double underlined. The stop codon is underlined with a wavy line.  
                                     (SEQ ID NO: 9)                  ATGAGGCAGTCTCTCCTATTCCTGACCAGCGTGGTTCCTTTCGTGCTGGC                       G   C   C   G   C   G   A   C   C   T   C   C   G   G   A   T   G   A   C   C   C   G   G   G   C                   T   T   C   G   G   C   C   C   C   C   A   C   C   A   G   A   G   G   C   T   C   G   A   G   A                   A   G   C   T   T   G   A   T   T   C   T   T   T   G   C   T   C   T   C   A   G   A   C   T   A                   C   G   A   T   A   T   T   C   T   C   T   C   T   T   T   A   T   C   T   A   A   T   A   T   C                   C   A   G   C   A   G   C   A   T   T   C   G   G   T   A   A   G   A   A   A   A   A   G   A   G                   A   T   C   T   A   C   A   G   A   C   T   T   C   A   A   C   A   C   A   T   G   T   A   G   A                   A   A   C   A   C   T   A   C   T   A   A   C   T   T   T   T   T   C   A   G   C   T   T   T   G                   A   A   A   A   G   G   C   A   T   T   T   T   A   A   A   T   T   A   T   A   C   C   T   G   A                   C   A   T   C   A   A   G   T   A   C   T   G   A   A   C   G   T   T   T   T   T   C   A   C   A                   A   A   A   T   T   T   C   A   A   G   G   T   C   G   T   G   G   T   G   G   T   G   G   A   T                   G   G   T   A   A   A   A   A   C   G   A   A   A   G   C   G   A   G   T   A   C   A   C   T   G                   T   A   A   A   A   T   G   G   C   A   G   G   A   C   T   T   C   T   T   C   A   C   T   G   G                   A   C   A   C   G   T   G   G   T   T   G   G   T   G   A   G   C   C   T   G   A   C   T   C   T                   A   G   G   G   T   T   C   T   A   G   C   C   C   a   C   A   T   A   A   G   A   G   A   T   G                   A   T   G   A   T   G   T   T   A   T   A   A   T   C   A   G   A   A   T   C   A   A   C   A   C                   A   G   A   T   G   G   G   G   C   C   G   A   A   T   A   T   A   A   C   A   T   A   G   A   G                   C   C   A   C   T   T   T   G   G   A   G   A   T   T   T   G   T   T   A   A   T   G   A   T   A                   C   C   A   A   A   G   A   C   A   A   A   A   G   A   A   T   G   T   T   A   G   T   T   T   A                   T   A   A   A   T   C   T   G   A   A   G   A   T   A   T   C   A   A   G   A   A   T   G   T   T                   T   C   A   C   G   T   T   T   G   C   A   G   T   C   T   C   C   A   A   A   A   G   T   G   T                   G   T   G   G   T   T   A   T   T   T   A   A   A   A   G   T   G   G   A   T   A   A   T   G   A                   A   G   A   G   T   T   G   C   T   C   C   C   A   A   A   A   G   G   G   T   T   A   G   T   A                   G   A   C   A   G   A   G   A   A   C   C   A   C   C   T   G   A   A   G   A   G   C   T   T   G                   T   T   C   A   T   C   G   A   G   T   G   A   A   A   A   G   A  AGAGCTGA               CCCAGATCCCATGAAGAACACGTGTAAATTATTGGTGGTAGCAGATCATC               GCTTCTACAGATACATGGGCAGAGGGGAAGAGAGTACAACTACAAATTAC               TTAATAGAGCTAATTGACAGAGTTGATGACATCTATCGGAACACTGCATG               GGATAATGCAGGTTTTAAAGGCTATGGAATACAGATAGAGCAGATTCGCA               TTCTCAAGTCTCCACAAGAGGTAAAACCTGGTGAAAAGCACTACAACATG               GCAAAAAGTTACCCAAATGAAGAAAAGGATGCTTGGGATGTGAAGATGTT               GCTAGAGCAATTTAGCTTTGATATAGCTGAGGAAGCATCTAAAGTTTGCT               TGGCACACCTTTTCACATACCAAGATTTTGATATGGGAAcTCtTGGATTA               GCTTATGTTGGCTCTCCCAGAGCAAACAGCCATGGAGGTGTTTGTCCAAA               GGCTTATTATAGCCCAGTTGGGAAGAAAAATATCTATTTGAATAGTGGTT               TGACGAGCACAAAGAATTATGGTAAAACCATCCTTACAAAGGAAGCTGAC               CTGGTTACAACTCATGAATTGGGACATAATTTTGGAGCAGAACATGATCC               GGATCGTCTAGCAGAATGTGCCCCGAATGAGGACCAGGGAGGGAAATATG               TCATGTATCCCATAGCTGTGAGTGGCGATCACGAGAACAATAAGATGTTT               TCACAGTGCAGTAAACAATCAATCTATAAGACCATTGAAAGTAAGGCCCA               GGAGTGTTTTCAAGAACGCAGCAATAAAGTT GGGAGCCACCATCATCACC                   ATCAC  {tilde under (TAA)}          
 
         [0077]     Any technique for mutagenesis known in the art can be used to convert the native TACE catalytic domain to a modified domain, including but not limited to, in vitro site-directed mutagenesis [Hutchinson et al.,  J. Biol. Chem.,  253:6551 (1978); Zoller and Smith,  DNA,  3:479-488 (1984); Oliphant et al.,  Gene,  44:177 (1986); Hutchinson et al.,  Proc. Natl. Acad. Sci. U.S.A.,  83:710 (1986)]. The use of TAB@ linkers (Pharmacia), etc. and PCR techniques also can be employed for site directed mutagenesis [see Higuchi, “Using PCR to Engineer DNA”, in PCR Technology: Principles and Applications for DNA Amplification, H. Erlich, ed., Stockton Press, Chapter 6, pp. 61-70 (1989)]. Preferably mutagenesis (i.e., modification) of the TACE catalytic domain is performed in a two step process [Wang, and Malcolm,  BioTechniques  26:680-682 (1999)]. In the Example below, two extension reactions were performed in separate tubes in the first stage: (i) one containing the forward primer, and (ii) the other containing the reverse primer. After two cycles, the two reactions are mixed and the standard QuickChange mutagenisis procedure is carried out for an additional 18 cycles. Following amplification, the parental strand is digested with lUnit of Dpn1 for 1 hour and an aliquot is transformed into DH5-alpha cells [GeneWiz, New York, N.Y.] 
       The TACE Polypeptide  
       [0078]     The amino acid sequence for the TACE polypeptide is shown below. (i) The pre domain is underlined, (ii) the pro domain is under-dashed, and (iii) the catalytic domain is unmarked. The GS-linker and polyhistidine tag (H6) are not included. The valine residue that is replaced with a non-hydrophobic amino acid residue in the modified TACE polypeptide is in bold. In addition, serine-266 has been replaced by an alanine, and asparagine-452 has been replaced by a glutamine in order to remove the N-linked glycosylation sites.  
                                     (SEQ ID NO: 2)                  MRQSLLFLTSVVPFVLA   P   R   P   P   D   D   P   G   F   G   P   H   Q   R   L   E   K   L   D   S   L   L                       S   D   Y   D   I   L   S   L   S   N   T                   Q   Q   H   S   V   R   K   R   D   L   Q   T   S   T   H   V   E   T   L   L   T   F   S   A   L                   K   R   H   F   K   L   Y   L   T   S   S   T   E   R   F   S   Q   N   F   K   V   V   V   V   D                   G   K   N   E   S   E   Y   T   V   K   W   Q   D   F   F   T   G   R   V   V   G   E   P   D   S                   R   V   L   A   H   I   R   D   D   D   V   I   I   R   I   N   T   D   G   A   E   Y   N   I   E                   P   L   W   R   F   V   N   D   T   K   D   K   R   M   L   V   Y   K   S   E   D   I   K   N   V                   S   R   L   Q   S   P   K   V   C   G   Y   L   K   V   D   N   E   E   L   L   P   K   G   L   V                   D   R   E   P   P   E   E   L   V   H   R   V   K   R RADPDPMKNTCKLLVVADHRFYRYMGRGEESTTTNY               LIELIDRVDDIYRNTAWDNAGFKGYGIQIEQIRILKSPQEVKPGEKHYNM               AKSYPNEEKDAWDVKMLLEQFSFDIAEEASKVCLAHLFTYQDFDMGTLGL               AY V GSPRANSHGGVCPKAYYSPVGKKNIYLNSGLTSTKNYGKTILTKEAD               LVTTHELGHNFGAEHDPDGLAECAPNEDQGGKYVMYPIAVSGDHENNKMF               SQCSKQSIYKTTESKAQECFQERSNKV          
 
         [0079]     The amino acid sequence for the catalytic domain of the modified TACE polypeptide is shown below. Whereas, the native protein comprises VAL 353  (VAL 139  of SEQ ID NO: 6), this amino acid residue is replaced by a non-hydrophobic amino acid residue in a modified TACE catalytic domain. This amino acid position is denoted with an “X” in bold below. Preferably the non-hydrophobic amino acid residue is a glycl, alanyl, or seryl amino acid residue. The GS-linker and Polyhistidine tag (H6) are not included.  
         [0080]     RADPDPMKNTCKLLVVADHRFYRYMGRGEESTTTNYLIELIDRVDDIYRNTAWDNAGFKG YGIQIEQIRILKSPQEVKPGEKHYNMAKSYPNEEKDAWDVKMLLEQFSFDIAEEASKVCL AHLFTYQDFDMGTLGLAYXGSPRANSHGGVCPKAYYSPVGKKNIYLNSGLTSTKNYGKTI LTKEADLVTTHELGHNFGAEHDPDGLAECAPNEDQGGKYVMYPIAVSGDHENNKMFSQCS KQSIYKTIESKAQECFQERSNKV (SEQ ID NO: 8), where X is a non-hydrophobic amino acid residue, and preferably either alanine, glycine or serine.  
         [0081]     In a particular embodiment of the present invention, a modified TACE polypeptide or fragment thereof (e.g., the catalytic domain) is at least about 75% identical, more preferably at least about 90% identical, and most preferably at least about 95% identical to the TACE polypeptide or fragment thereof. As indicated above, a modified TACE or fragment thereof has a non-hydrophobic amino acid residue in place of the valine at position 353 (as defined in SEQ ID NO: 2).  
         [0082]     Polypeptides comprising the modified TACE catalytic domains of the invention include those containing altered sequences in which functionally equivalent amino acid residues are substituted for residues within the sequence resulting in a conservative amino acid substitution. For example, one or more amino acid residues within the sequence can be substituted by another amino acid of a similar polarity, which acts as a functional equivalent, resulting in a silent alteration. Substitutes for an amino acid within the sequence may be selected from other members of the class to which the amino acid belongs.  
         [0083]     For example, the nonpolar amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. Amino acids containing aromatic ring structures are phenylalanine, tryptophan, and tyrosine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. The positively charged (basic) amino acids include arginine and lysine. The negatively charged (acidic) amino acids include aspartic acid and glutamic acid.  
         [0084]     Particularly preferred conserved amino acid exchanges are:  
         [0085]     (a) Lys for Arg or vice versa such that a positive charge may be maintained;  
         [0086]     (b) Glu for Asp or vice versa such that a negative charge may be maintained;  
         [0087]     (c) Ser for Thr or vice versa such that a free —OH can be maintained;  
         [0088]     (d) Gln for Asn or vice versa such that a free NH 2  can be maintained; and  
         [0089]     (e) Ile for Leu or for Val or vice versa as roughly equivalent hydrophobic amino acids.  
         [0090]     All of the modified TACE catalytic domains of the present invention also can be part of a chimeric protein. In a specific embodiment, a chimeric TACE protein is expressed in a eukaryotic cell. Such a chimeric protein can be a fusion protein used to isolate a modified TACE of the present invention, through the use of an affinity column that is specific for the protein fused to the TACE protein. In one such embodiment, the chimeric TACE is expressed in a eukaryotic cell. Examples of such fusion proteins include: a glutathione-S-transferase (GST) fusion protein, a maltose-binding (MBP) protein fusion protein, a FLAG-tagged fusion protein, or as specifically exemplified below, a poly-histidine-tagged fusion protein.  
         [0091]     Expression of a chimeric TACE protein, or fragment thereof, as a fusion protein can facilitate stable expression, and/or allow for purification based on the properties of the fusion partner. Thus the purification of the recombinant polypeptides of the present invention can be simplified through the use of fusion proteins having affinity tags. For example, GST binds glutathione conjugated to a solid support matrix, MBP binds to a maltose matrix, and poly-histidine chelates to a NI-chelation support matrix, as specifically exemplified below [see Hochuli et al.,  Biotechnolgy  6:1321-1325 (1998)]. The fusion protein can be eluted from the specific matrix with appropriate buffers, or by treating with a protease that is specific for a cleavage site that has been genetically engineered in between the TACE protein and its fusion partner. Alternatively, a modified TACE catalytic domain can be combined with a marker protein such as green fluorescent protein [Waldo et al.,  Nature Biotech.  17:691-695 (1999); U.S. Pat. No. 5,625,048 filed Apr. 29, 1997 and WO 97/26333, published Jul. 24, 1997, the contents of which are hereby incorporated by reference herein in their entireties].  
         [0092]     Alternatively or in addition, other column chromatography steps (e.g., gel filtration, ion exchange, affinity chromatography etc.) can be used to purify the recombinant proteins of the present invention. In many cases, such column chromatography steps employ high performance liquid chromatography or analogous methods in place of the more classical gravity-based procedures.  
         [0093]     As exemplified below, a recombinant modified TACE catalytic domain was purified with a NiNTA column following routine centrifugation and diafiltration steps. After the purified protein was collected from the NiNTA column, it was placed on a gel filtration column. The resulting eluate was then concentrated and desalted prior to being combined with an inhibitor to form a protein-ligand complex.  
         [0094]     Alternatively, polypeptides comprising the modified TACE catalytic domains of the present invention can be chemically synthesized [see e.g., Synthetic Peptides:  A User&#39;s Guide , W.H. Freeman &amp; Co., New York, N.Y., pp. 382, Grant, ed. (1992)].  
       Enzyme Assays  
       [0095]     The catalytic activity of the TACE protease can be determined in an assay using the synthetic peptide Ac-SPLAQA-VRSSSR-NH 2  (SEQ ID NO: 17) as the substrate. This amino acid sequence corresponds to the cleavage site of TACE on proTNF-alpha, with the sessile bond being between the alanine and the valine. The activity can be measured by incubating 100 nM TACE with 100 micromolar substrate in 25 mM HEPES pH 7.3, 5 mM CaCl 2 , for 1 hour at room temperature. Product formation can be quantified at 214 nm by HPLC using a reverse phase column to separate the substrate from the products. The ability of a given compound added to the reaction to act as an inhibitor of TACE can then be determined.  
         [0096]     Alternatively, TACE activity can be determined in a fluorescence assay using the synthetic peptide substrate, K(Mca)-SPLAQA-VRSSSRK(Dpn)-NH 2  (SEQ ID NO: 18). K(Mca) is a lysyl residue modified by comprising an epsilonN-methoxycoumarin, whereas K(Dpn)-NH 2  is a lysyl residue modified to comprise an epsilonN 2,4, dinitrophenyl. 2-100 nanomolar TACE protease (or active fragment thereof) is incubated with 25 micromolar peptide substrate in 25 mM HEPES pH 7.3, 5 mM CaCl 2  for 1 hour at room temperature. Product formation is detected by exciting at 340 nm and measuring the fluorescence emission at 380 nm every 30 seconds for about an hour. The initial velocity can be obtained by linear regression. The increase in fluorescence emission can be correlated with the quantity of cleaved product. The ability of a given compound added to the reaction to act as an inhibitor of TACE can then be determined.  
       Crystallization  
       [0097]     Crystals of the protein-ligand complex comprising a modified TACE catalytic domain of the present invention can be grown by a number of techniques including batch crystallization, vapor diffusion (e.g., by sitting drop or hanging drop) and by microdialysis. In the Example below, the modified TACE catalytic domain was complexed with N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide and crystallized by hanging drop vapor diffusion. Seeding of the crystals in some instances is required to obtain X-ray quality crystals. Standard micro and/or macro seeding of crystals may therefore be used.  
         [0098]     As exemplified below, the protein-ligand complex comprising the modified TACE catalytic domain V353G (vgTACE) was crystallized under similar conditions to those previously employed for the non-modified TACE [WO9940182, Published Aug. 12, 1999, U.S. application Ser. No. 09/117,476, filed Jan. 27, 1999, the contents of which are both hereby incorporated by reference in its entireties].  
         [0099]     A substitute ligand can replace the co-crystallized initial ligand by soaking a crystal of protein-initial ligand complex with the substitute ligand. Thus, one or more crystals of protein-initial ligand complex can be placed in the reservoir solution containing about a 10-fold or greater excess of substitute ligand. The crystal is kept under the appropriate conditions and for a sufficient time period for the substitute ligand to replace the initial ligand and form the new crystalline protein-substitute ligand complex. In the example below, a crystal was kept in the solution containing the substitute ligand for about 72 hours. After the incubation, the crystal of the protein-substitute ligand complex can be frozen in liquid propane, for example and then used for X-ray diffraction.  
         [0100]     Crystals can be characterized using X-rays produced in a conventional source (such as a sealed tube or a rotating anode) or using a synchrotron source. Methods of characterization include, but are not limited to, precision photography, oscillation photography and diffractometer data collection. As exemplified below, the crystals were flash frozen in liquid propane and X-ray diffraction was collected at 100 degrees Kelvin using conventional or synchrotron sources.  
         [0101]     In the Example below, the crystal structure of the modified TACE catalytic domain V353G (vgTACE) was solved by molecular replacement and then refined using standard crystallographic programs. The published TACE structure was used as the starting model [PDB code: 1BKC; Maskos et. al.,  Proc. Natl. Acad. Sci. USA  95:3408-3412 (1998); WO9940182, Published Aug. 12, 1999, U.S. application Ser. No. 09/117,476, filed Jan. 27, 1999, the contents of which are both hereby incorporated by reference in its entirety]. Replacement of the co-crystallized inhibitor was verified by difference electron density maps. The vgTACE:inhibitor structures were refined using X-PLOR [Brunger et al.,  Acta Crystallogr . A 46:585-593 (1990); Brunger et al.,  Acta Crystallogr. D Biol. Crystallogr.,  54:905-921 (1998)].  
         [0102]     Refinement calculations also can be performed using CNS [Adams et al.,  Proc. Natl. Acad. Sci. USA,  94:5018-5023 (1997)]. Map interpretation and model building also can be performed using O [Jones et al.,  Acta Cryst , A 47:110-119 (1991)]. Other computer programs that can be used to solve crystal structures include: QUANTA, CHARMM; INSIGHT; SYBYL; MACROMODE; and ICM.  
         [0103]     Generally, structure based rational drug design is performed by analyzing the three-dimensional structures of successive protein-ligand complexes. This iterative process requires X-ray quality crystals of numerous protein-ligand complexes. These crystals can be obtained three ways. First, crystals of each protein-ligand complex can be grown de novo. This is the most time-consuming method, and in many instances requires determining a new set of crystallization conditions. The second method is to incubate (e.g., soak) individual crystals of the uncomplexed protein with each different ligand. This method is much faster than growing new crystals, but still requires a relatively large stock of protein to generate all of the new crystals. The third and most expedient method is to incubate a previously formed protein-ligand crystal with a large excess of a substitute ligand, thereby replacing the initial ligand with the substitute ligand in the protein-ligand complex. Heretofore, it was difficult to prepare alternative protein-ligand complexes of TACE since the two available X-ray quality crystals of TACE comprised the unstable, native TACE. The present invention overcomes this problem by providing a modified TACE catalytic domain that forms X-ray quality crystals that are amenable to ligand exchange.  
       Structure Based Rational Drug Design  
       [0104]     Once three-dimensional structures of crystals comprising modified TACE catalytic domains are determined, a potential inhibitor of TACE can be examined through the use of computer modeling using a docking program such as GRAM, DOCK, or AUTODOCK [Dunbrack et al.,  Folding &amp; Design,  2:27-42 (1997)]. This procedure can include computer fitting of potential inhibitors to the modified TACE catalytic domain to ascertain how well the shape and the chemical structure of the potential modulator will interact with the TACE protein [Bugg et al.,  Scientific American , Dec.:92-98 (1993); West et al.,  TIBS,  16:67-74 (1995)]. Computer programs can also be employed to estimate the attraction, repulsion, and steric hindrance of the modified TACE catalytic domain with an inhibitor.  
         [0105]     Generally the tighter the fit, the lower the steric hindrances, and the greater the attractive forces, the more potent the inhibitor, since these properties are consistent with a tighter binding constant. Furthermore, the more specificity in the design of a potential drug the more likely that the drug will not interact as well with other proteins. This will minimize potential side-effects due to unwanted interactions with other proteins.  
         [0106]     Initially compounds known to bind TACE, for example N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide, or a compound that inhibits TACE disclosed by Letavic et al., [ Biorgan. &amp; Medic. Chem Lett.  12:1387-1390 (2002) the contents of which are hereby incorporated by reference in their entireties], or alternatively, a compound that binds metalloproteases as disclosed as by Zask et al. [ Curr. Pharm. Des.,  2:624-661 (1996), the contents of which are hereby incorporated by reference in their entireties], can be systematically modified by computer modeling programs until one or more promising potential analogs are identified. Such analysis has been shown to be effective in the development of HIV protease inhibitors [Lam et al.,  Science  263:380-384 (1994); Wlodawer et al.,  Ann. Rev. Biochem.  62:543-585 (1993); Appelt,  Perspectives in Drug Discovery and Design  1:23-48 (1993); Erickson, Perspectives in Drug Discovery and Design 1:109-128 (1993)]. Alternatively, a potential inhibitor initially can be obtained by screening a random peptide library or a chemical library. In the former case, a random peptide library can be produced by recombinant bacteriophage, for example, [Scott and Smith,  Science,  249:386-390 (1990); Cwirla et al.,  Proc. Natl. Acad. Sci.,  87:6378-6382 (1990); Devlin et al.,  Science,  249:404-406 (1990)]. A peptide selected in this manner would then be systematically modified by computer modeling programs, as described above.  
         [0107]     If a potential inhibitor is a small organic compound, it can be selected from a library of chemicals, including commercially available chemical libraries. Alternatively, the small organic compound may be synthesized de novo. The de novo synthesis of one or even a relatively small group of specific compounds is reasonable in the art of drug design. Once obtained, the potential inhibitor can be further tested in a standard binding and/or catalytic assay with TACE, the TACE catalytic domain, or an active fragment thereof.  
         [0108]     For example, a binding assay can be performed following the attachment of the TACE catalytic domain to a solid support. Methods for placing the TACE catalytic domain on the solid support are well known in the art and include such things as linking biotin to the TACE catalytic domain and linking avidin to the solid support. The solid support can be washed to remove unbound protein. A solution of a labeled potential inhibitor can be contacted with the solid support. The solid support is washed again to remove the potential inhibitor not bound to the support. The amount of labeled potential inhibitor remaining with the solid support, and thereby bound to the TACE catalytic domain can be determined. Alternatively, or in addition, the dissociation constant between the labeled potential inhibitor and the TACE catalytic domain, for example, can be determined.  
         [0109]     Suitable labels for either the TACE catalytic domain or the potential inhibitor include, radioactive labels (e.g.,  14 C,  1 H,) and fluorescent labels such as fluorescein isothiocyanate (FITC).  
         [0110]     In another embodiment, a Biacore machine can be used to determine the binding constant of the TACE catalytic domain with a potential inhibitor [O&#39;Shannessy et al.  Anal. Biochem.  212:457-468 (1993); Schuster et al.,  Nature  365:343-347 (1993)]. In another aspect of the present invention a potential inhibitor is tested for its ability to inhibit the proteolytic activity of TACE or an active fragment thereof. An inhibitor is then selected on the basis of its ability to inhibit the catalytic reaction of the TACE protease.  
         [0111]     When a promising inhibitor is identified, a crystal comprising a protein-ligand complex of the inhibitor and the modified TACE catalytic domain can be prepared by incubating an excess of the inhibitor (substitute ligand) with a crystal of a modified TACE catalytic domain-ligand complex. The three-dimensional structure of the resulting crystalline protein-substitute ligand complex can then be determined by molecular replacement analysis, for example.  
         [0112]     Molecular replacement involves using a known three-dimensional structure as a search model to determine the structure of a closely related molecule or protein-ligand complex in a different crystalline form. The measured X-ray diffraction properties of the new crystal are compared with the search model structure to compute the position and orientation of the protein in the new crystal. Computer programs that can be used include: X-PLOR (see above), CNS, (Crystallography and NMR System, a next level of XPLOR), and AMORE [Navaza,  Acta Crystallographics  ASO, 157-163 (1994)]. Once the position and orientation are known, an electron density map can be calculated using the search model to provide X-ray phases. Thereafter, the electron density is inspected for structural differences and the search model is modified to conform to the new structure. Using this approach, it is possible to solve the three-dimensional structures of crystals of any protein-ligand complex of the modified TACE catalytic domain.  
         [0113]     For all of the drug screening assays described herein, further refinements to the structure of the drug will generally be necessary and can be made by the successive iterations of any and/or all of the steps provided by the particular drug screening assay and/or in combination with other such drug screening assays.  
         [0114]     A candidate drug selected by performing structure based rational drug design can then be assayed in situ and/or in vivo. A candidate drug can be identified as a drug, for example, if it ameliorates a symptom caused by an overabundance of the soluble form of TNF-alpha in an animal model. Indeed, methods of testing such potential candidate drugs in animal models are well known in the art. The potential drugs can be administered by a variety of ways including topically, orally, subcutaneously, or intraperitoneally depending on the proposed use. Generally, at least two groups of animals are used in the assay, with at least one group being a control group that is administered the administration vehicle without the potential drug.  
       Electronic Representation of the Three Dimensional Structure of TACE  
       [0115]     The present invention provides the three-dimensional depiction of the TACE catalytic domain in a complex with an inhibitor on an electronic and/or magnetic medium. More specifically, the present invention provides the data comprised in Table 3 on an electronic and/or magnetic medium. In addition, the present invention provides a computer that comprises a representation of the TACE catalytic domain-inhibitor complex in computer memory that can be used to screen for compounds that will inhibit the proteolytic activity of TACE. The computer may comprise portions of, or all of the information contained in Table 3. In a particular embodiment, the computer comprises: (i) a machine-readable data storage material encoded with machine-readable data, (ii) a working memory for storing instructions for processing the machine readable data, (iii) a central processing unit coupled to the working memory and the machine-readable data storage material for processing the machine readable data into a three-dimensional representation, and (iv) a display coupled to the central processing unit for displaying the three-dimensional representation. Thus the machine-readable data storage medium comprises a data storage material encoded with machine readable data which can comprise portions of, or all of the structural information contained in Table 3.  
         [0116]     One embodiment for manipulating and displaying the structural data provided by the present invention is schematically depicted in  FIG. 1 . As depicted the System 1, includes a computer  2  comprising a central processing unit (“CPU”)  3 , a working memory  4  which may be random-access memory or “core” memory, mass storage memory  5  (e.g., one or more disk or CD-ROM drives), a display terminal  6  (e.g., a cathoderay tube), one or more keyboards  7 , one or more input lines  10 , and one or more output lines  20 , all of which are interconnected by a conventional bidirectional system bus  30 .  
         [0117]     Input hardware  12 , coupled to the computer  2  by input lines  10 , may be implemented in a variety of ways. Machine-readable data may be inputted via the use of one or more modems  14  connected by a telephone line or dedicated data line  16 . Alternatively or additionally, the input hardware may comprise CD-ROM or disk drives  5 . In conjunction with the display terminal  6 , the keyboard  7  may also be used as an input device. Output hardware  22 , coupled to computer  2  by output lines  20 , may similarly be implemented by conventional devices. Output hardware  22  may include a display terminal  6  for displaying the three dimensional data. Output hardware might also include a printer  24 , so that a hard copy output may be produced, or a disk drive or CDROM  5 , to store system output for later use, [see also U.S. Pat. No. 5,978,740, Issued Nov. 2, 1999, the contents of which are hereby incorporated by reference in their entireties].  
         [0118]     In operation, the CPU  3  (i) coordinates the use of the various input and output devices  12  and  22 ; (ii) coordinates data accesses from mass storage  5  and accesses to and from working memory  4 ; and (iii) determines the sequence of data processing steps. Any of a number of programs may be used to process the machine-readable data of this invention.  
                                           TABLE 1                           TABLE OF SEQUENCES            SEQ ID NO:   Type   Description                    1   N.A.   Pre, Pro, and Catalytic domain       2   A.A.   Pre, Pro, and Catalytic domain       3   N.A.   Pro and Catalytic domain       4   A.A.   Pro, and Catalytic domain       5   N.A.   Catalytic domain       6   A.A.   Catalytic domain       7   N.A.   Catalytic domain (modified)       8   A.A.   Catalytic domain (modified)       9   N.A.   open reading frame       10   N.A.   BamH1f primer       11   N.A.   Kpn1r primer       12   N.A.   V353Gf primer       13   N.A.   V353Gr primer       14   N.A.   V353Sf primer       15   N.A.   V353Sr primer       16   A.A   internal cleave site       17   A.A   synthetic TACE substrate       18   A.A   synthetic TACE substrate       19   N.A.   vgTACE       20   A.A   vgTACE                  
 
         [0119]     The present invention may be better understood by reference to the following non-limiting Example, which is provided as exemplary of the invention. The following example is presented in order to more fully illustrate the preferred embodiments of the invention. It should in no way be construed, however, as limiting the broad scope of the invention.  
       EXAMPLE  
     Material and Methods  
       [0000]     Cloning of TACE and TACE Mutants:  
         [0120]     The TACE protein was cloned and expressed as the pro-protein. The pre and pro domains are cleaved during protein expression and secretion by the cells. Only the catalytic domain was purified.  
         [0121]     Two PCR primers were used to amplify the pre-pro-cat domains of TACE having BamH1 and GS-(His)6-Kpn1 sites at the 5′ and 3′ ends respectively:  
                                                                 SEQ ID NO: 10                BamH1f:   5′cgcggatccatgaggcagtctctcctattcctg 3′                        SEQ ID NO: 11                Kpn1r:   5′ccggcctaccttagtgatggtgatgatggtgggatc 3′              
 
         [0122]     The purified PCR fragment was digested with BamH1 and Kpn1 and subcoloned into the pFastBac1 vector provided in the Bac-to-Bac Baculovirus expression system (Invitrogen, Carlsbad, Calif.). The TACE mutants (V353G, V353S) were generated using the QUICKCHANGE kit (Stratagene, La Jolla, Calif., USA) using the native TACE pFastBac1 vector as a template and the following complementary mutagenic primers:  
                               V353Gf:               5′GGA ACT CTT GGA TTA GCT TAT GGA GGC   SEQ ID NO: 12               TCT CCC AGA GCA AAC 3′               V353Gr:       5′GTT TGC TCT GGG AGA GCC TCC ATA AGC   SEQ ID NO: 13               TAA TCC AAG AGT TCC 3′               V353Sf:       5′GGA ACT CTT GGA TTA GCT TAT AGC GGC   SEQ ID NO: 14               TCT CCC AGA GCA AAC3′               V353Sr:       5′GTT TGC TCT GGG AGA GCC GCT ATA AGC   SEQ ID NO: 15               TAA TCC AAG AGT TCC3′          
 
         [0123]     The mutagenesis was performed in two steps as previously described [Wang, and Malcolm,  BioTechniques  26:680-682 (1999) the contents of which are hereby incorporated by reference in their entireties]. In the first stage, two extension reactions were performed in separate tubes; one containing the forward primer and the other containing the reverse primer. After two cycles, the two reactions were mixed and the standard QUICKCHANGE mutagenisis procedure was carried out for an additional 18 cycles. Following amplification, the parental strand was digested with 1Unit of Dpn1 for 1 hour and an aliquot was transformed into DH5-alpha cells. The sequences of all of the vectors were confirmed. (GeneWiz, New York, N.Y.)  
         [0124]     Production of Recombinant Baculovirus: Recombinant baculovirus was produced using the BAC-TO-BAC expression system (Invitrogen, Carlsbad, Calif.) following known protocols for the transposition, isolation and transfection of recombinant bacmid DNA into Sf9 cells for production of viral particles. The virus was amplified to the P2 generation and was titered using the BacPAC Baculovirus Rapid Titer Kit (Clonetech, Palo Alto, Calif.).  
         [0125]     Expression and Purification of TACE and TACE mutants: Logarithmically growing Trichoplusia Ni cells (High-5 TM  cells, 2×10 6  cells/ml) were infected with amplified baculovirus at a MOI=1.0 (2.5×10 8  pfu/ml) and grown at 27 degrees Celsius for 48-60 hours. Secreted TACE was isolated from the cell culture media after clarification by centrifugation. The pooled supernatants were concentrated 10 fold and the buffer exchanged into 25 mM HEPES, 0.15M NaCl, pH 7.5 by diafiltration. To the desalted supernatant, 4-aminophenyl-mercuric acetate (APMA) was added to 20 μM, lauryl maltoside to 0.05%, and imidazole to 25 mM. The supernatant was then applied to a NiNTA column (Qiagen Hilden, Germany). The NiNTA column was washed with 25 mM imidazole in buffer A (50 mM HEPES, 10% glycerol, 0.3M NaCl, 0.1% m-octyl-Beta-D-glucopyranoside, pH 7.5) until a stable baseline was achieved. The protein was then eluted with 250 mM imidazole in buffer A. The eluted protein was diluted to 0.1 mg/ml and dialyzed overnight against 25 mM Tris pH 7.5, with 20 μM APMA to digest excess pro-domain. The protein was collected and adjusted to 0.15M NaCl, concentrated, and applied to a SUPERDEX-75 gel filtration column (Pharmacia) equilibrated with 25 mM Tris-HCl, 0.2M NaCl pH 7.5 at 4 degrees Celsius. Fractions corresponding to the monomer of TACE were pooled, and stored at 4 degrees Celsius. The pooled TACE enzyme was concentrated to 15 mg/ml, desalted into 25 mM Tris-HCl pH 7.5 using BIO-SPIN 6 columns (Bio-Rad, Hercules, Calif.) and immediately complexed with N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide at a 1:1.5 molar ratio. Using this protocol, total expression levels of 5-10 mg/L were obtained with a final recovery of 0.5-5 mg/L.  
         [0126]     Crystallization: Crystals were obtained by the hanging drop vapor diffusion method [Ducruix and Giege.  Crystallization of Nucleic Acids and Proteins . A practical approach. Oxford University Press, (1992)]. A small volume (1 to 15 microliters) containing the N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl }-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide [commercially available from e.g., CALBIOCHEM, San Diego Calif., Catalogue No. 579052, (TAPI-2)] solution was equilibrated with a larger volume (1 ml) of a reservoir solution. The reservoir solution contains the precipitant that facilitates the crystallization.  
         [0127]     During equilibration the water content in the hanging drop is reduced and the protein-ligand complex, [i.e., the complex between vgTACE and N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide] forms crystals. To crystallize the protein-ligand complex, one microliter of the protein-ligand complex solution was mixed with one microliter of the reservoir solution, which contains 15% polyethylenglycol 4000, 10% 2-Propanol, and 100 mM Citrate-Buffer pH 5.6. Crystals were observed after one week.  
         [0128]     The crystals obtained were washed with the reservoir solution. In the next step, a single protein-ligand complex crystal was put into the reservoir solution, which contained in addition, a 10 mM TACE inhibitor to replace the co-crystallized N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide. The crystal is usually kept for 72 hours in the solution containing the inhibitor to allow the ligand replacement. After incubation, the crystal was frozen in liquid propane and used for X-ray diffraction.  
       Results  
       [0129]     Two distinct structures of TACE:inhibitor complexes have been previously disclosed [Maskos et. al.,  Proc. Natl. Acad. Sci. USA  95:3408-3412 (1998); Letavic et al.  Biorgan. &amp; Medic. Chem Lett.  12:1387-1390 (2002)]. However, neither crystal appears to be amenable to crystal soaking. As disclosed herein, it has been unexpectedly discovered that the uncomplexed TACE protein (apo-protein) is unstable at the high concentrations required to grow and use crystals for X-ray crystallographic studies. Consistently, the crystal of Maskos et. al. has been found to be resistant to standard inhibitor soaking experiments, severely limiting its value in structure based rational drug design.  
         [0130]     Stability of TACE: The stability of TACE was examined under several buffer conditions at pH 7.5. Twenty microliter aliquots of TACE at 15 mg/ml were desalted over P-6 spin columns (BioRAD, Hercules, Calif.) that had been equilibrated in: 
        (a) 25 mM Tris, 0.15M NaCl;     (b) 25 mM Tris; or     (c) 25 mM Tris plus N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide.        
 
         [0134]     The stability of the TACE protein was evaluated after storage at 4 degrees Celsius for seven days. After incubating the TACE polypeptide under the three conditions listed above, Sodium Dodecyl Sulfate PolyAcrylamide Gel Electrophoresis (SDS-PAGE) was performed. The results show that only a single band having a molecular weight of 30 Kd was observed when either high salt (0.15 sodium chloride) or an inhibitor [N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide, at a 1:1.5 molar ratio enzyme:inhibitor] was included in the incubation. In direct contrast, two additional fainter bands that ran well ahead of the more significant TACE band were observed in the sample lacking either high salt or the inhibitor. The molecular weights of these two additional fainter bands were 14 Kd and 16 Kd, respectively, which add up to the 30 Kd molecular weight of the TACE polypeptide. N-terminal sequencing of the peptides corresponding to the two additional bands indicated that they were indeed, proteolytic products of the TACE protein. Moreover, the sequencing data indicated the presence of a single cleavage site at 352Y-V353 of SEQ ID NO: 2.  
         [0135]     Substrate Specificity of TACE: In an effort to understand the role of the different amino-acid residues of TACE regarding substrate specificity, a substitution study was performed at the P′1 position. The catalytic activity of TACE was determined in an assay using the synthetic peptide Ac-SPLAQA-VRSSSR-NH2 (SEQ ID NO: 17) as the substrate. The sequence corresponds to the cleavage site of TACE on proTNF-alpha, with the sessile bond being between the alanine and the valine. Activity was measured by incubating 100 nM TACE with 100 micromolar substrate in 25 mM HEPES pH 7.3, 5 mM CaCl 2 , for 1 hour at room temperature. Product formation was quantified at 214 nm after HPLC separation using a POROS-R1 reverse phase column. Substitution of the P′1 valine (in bold above) with either alanine, glycine or serine decreased activity of TACE to non-detectable levels. Based on this data it was decided to substitute the P′1 position of the internal cleave site [ . . . LGLAY-VGSPR . . . (SEQ ID NO: 16)] with one of these amino acids e.g., either glycine or serine, in an attempt to eliminate the auto-proteolysis of TACE seen in the absence of NaCl.  
         [0136]     Stability of TACE and TACE mutants: To test for stability under different storage conditions, 20 ul of TACE protein, at 15 mg/ml, was desalted over BIO-RAD P-6 columns equilibrated at pH 7.5 in: 
        (a) 25 mM Tris, 0.15M NaCl;     (b) 25 mM Tris+1 mM N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide; or     (c) 25 mM Tris.        
 
         [0140]     One microliter aliquots were analyzed by SDS-PAGE after storing them for 3 hours, or 17 days at 4 degrees Celsius. In the absence of salt, the native protein exhibits a pattern consistent with substantial proteolysis occurring after only 3 hours, with the protein being completely proteolyzed after 17 days. In direct contrast, all constructs were stable in either 0.15M NaCl , or 1 mM N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide. The 2 loop mutants V353G and V353S showed improved stability, with the V353G (vgTACE) mutant being highly resistant to auto-proteolysis, even after 17 days.  
         [0141]     Crystallization: The TACE mutant V353G (vgTACE) could be crystallized under similar conditions as the native TACE [WO9940182, Published Aug. 12, 1999, U.S. Application No. 09/117,476, filed Jan. 27,1999, the contents of which are both hereby incorporated by reference in its entirety]. vgTACE was concentrated to 15 mg/ml in 150 mM NaCl, 25 mM Tris-HCl pH 8. After desalting the vgTACE with Bio-Rad P-6 columns, N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide was added to a molecular ratio of 1:2 (enzyme:inhibitor). The complex was crystallized using the hanging drop vapor diffusion technique. Equal amounts of vgTACE-inhibitor solution were mixed with the reservoir solution, containing 15% Polyethylenglycol 4000, 10% 2-propanol, 100 mM sodium citrate pH 4.6, and equilibrated at 295 degrees Kelvin. Crystals were observed after 7 days.  
         [0142]     Crystallographic analysis: vgTACE crystals were washed using the reservoir solution. Glycerol was then added to the reservoir solution to a final concentration of 15%, and the crystals were flash frozen in liquid propane. X-ray diffraction data were collected at 100 degrees Kelvin, using a rotating anode generator (Rigaku/MSC) or synchrotron sources. Diffraction was observed up to 1.7 A. The vgTACE crystals belong to space group P2 1 2 1 2 1 , (a=73, b=75, c=103 Å). There are two molecules located within the asymmetric unit. Soaking with the compounds of Table 2 was performed by incubation of the crystals in the reservoir solution in the presence of up to 70 mM of the respective inhibitor. In the V353G mutant crystals, the co-crystallized inhibitor N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide could be replaced during soaking.  
         [0143]     Model building and refinement: The crystal structure of V353G mutant was solved by molecular replacement and refined using standard crystallographic programs. The published TACE structure was used as the starting model [PDB code:1BKC; Maskos et. al.,  Proc. Natl. Acad. Sci. USA  95:3408-3412 (1998)]. Replacement of the co-crystallized inhibitor was verified by difference electron density maps. The vgTACE:inhibitor structures were refined using X-PLOR (CNS). A list of vgTACE:inhibitor structures that have been solved is shown in Table 2 below.  
                         TABLE 2                           RESOLUTION OF TACE-INHIBITOR COMPLEXES                Resolution       INHIBITOR   (Angstroms)               (4-biphenylsulfonyl)amino-propionic acid   1.7       3,4-dimethyl-2-(N-3′-pyridylmethyl-p-   1.9       methoxysulfonamido)-benzenehydroxyamic acid       alpha(R)-[[[4-2-butynyloxy) phenyl] sulfonyl] amino]   1.7       benzenepropanoic acid       N-hydroxy-alpha(R),3(S)-dimethyl-3-[4-[(2-methyl-4-   2.1       quinolinyl)methoxy]phenyl]-2-oxo-1-pyrrolidineacetamide       N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-   2.0       Ile-Leu-OH                  
 
         [0144]     Table 3 below, comprises the coordinate set from the crystal structure of TACE complexed with N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH. To obtain these coordinates a single co-crystal of TACE in complex with N-{D,L-2-(hydroxyaminocarbonyl)methyl-4-methylpentanoyl}-L-3-amino-2-dimethylbutanoyl-L-alanine, 2-(amino)ethyl amide was soaked in 10% polyethylenglycol 8000, 50 mM sodium citrate, 10 mM [N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH] and 10% dimethylsulphoxide (DMSO) for three days. The crystal was transferred into a solution with additional 10% glycerol and flash-frozen in liquid nitrogen. X-ray diffraction data were collected as described above. These data were processed and the structure was refined as described above.  
         [0145]     The TACE monomer includes amino acid residues 6-259 of the catalytic domain (see SEQ ID NO: 20) and is bound to the inhibitor, N-{3-(hydroxyaminocarbonyl)-1-oxo-(2R)-benzylpropyl}-Ile-Leu-OH which has the number 260 in Table 3. The catalytic zinc ion has the number 261. Amino acid residues Arg 28 , Lys 72 , Glu 81 , Lys 88 , Glu 93 , Glu 94 , Lys 95 , and Arg 143  were modeled as Ala residues since their side chains were disordered.  
         [0146]     The TACE protein used had the amino acid sequence of SEQ ID NO: 20, i.e., Gly 139  is the single point mutation site replacing Val 139  of the TACE wild-type amino acid sequence.  
         [0147]     In the data set below, one line contains information per one atom. The seven columns of Table 3 represent respectively: 
        1) residue number,     2) one-letter amino acid code,     3) atom name,     4) x-coordinate,     5) y-coordinate,     6) z-coordinate, and        
 
         [0154]     7) B-factor.  
                                             TABLE 3                       COORDINATES for the COMPLEX of vgTACE with       N-{3-(Hydroxyaminocarbonyl)-1-Oxo-(2R)-Benzylpropyl}-Ile-Leu-OH                                6   PRO   CB   62.073   53.680   60.089   57.01       6   PRO   CG   62.468   55.126   60.364   57.06       6   PRO   C   59.660   54.054   59.521   56.73       6   PRO   O   59.094   53.151   60.132   56.72       6   PRO   N   61.472   55.081   58.226   56.90       6   PRO   CD   62.545   55.773   58.963   57.18       6   PRO   CA   61.066   53.868   58.966   56.85       7   MET   N   59.104   55.240   59.307   56.47       7   MET   CA   57.761   55.534   59.770   56.16       7   MET   CB   57.653   57.002   60.180   57.02       7   MET   CG   58.514   57.370   61.375   58.06       7   MET   SD   58.165   59.050   61.936   59.40       7   MET   CE   59.102   59.999   60.715   59.24       7   MET   C   56.780   55.226   58.644   55.35       7   MET   O   55.612   54.925   58.885   55.64       8   LYS   N   57.269   55.306   57.411   54.05       8   LYS   CA   56.458   55.020   56.236   52.67       8   LYS   CB   56.923   55.885   55.062   53.02       8   LYS   CG   56.888   57.377   55.362   53.28       8   LYS   CD   57.228   58.210   54.135   53.51       8   LYS   CE   57.057   59.698   54.425   53.52       8   LYS   NZ   57.362   60.531   53.233   53.75       8   LYS   C   56.625   53.534   55.909   51.45       8   LYS   O   57.436   53.153   55.066   51.21       9   ASN   N   55.843   52.701   56.585   49.93       9   ASN   CA   55.927   51.263   56.398   48.31       9   ASN   CB   56.409   50.618   57.696   48.36       9   ASN   CG   55.496   50.928   58.878   48.21       9   ASN   OD1   55.835   50.643   60.029   48.47       9   ASN   ND2   54.333   51.504   58.599   48.06       9   ASN   C   54.610   50.632   55.987   47.19       9   ASN   O   54.456   49.415   56.074   47.11       10   THR   N   53.662   51.446   55.540   45.73       10   THR   CA   52.367   50.913   55.151   44.26       10   THR   CB   51.264   51.294   56.156   44.15       10   THR   OG1   51.681   50.975   57.487   44.15       10   THR   CG2   49.992   50.519   55.834   43.84       10   THR   C   51.883   51.361   53.787   43.31       10   THR   O   51.918   52.546   53.454   43.14       11   CYS   N   51.420   50.393   53.007   42.09       11   CYS   CA   50.872   50.663   51.693   41.05       11   CYS   C   49.373   50.626   51.927   40.79       11   CYS   O   48.813   49.571   52.249   40.43       11   CYS   CB   51.310   49.574   50.705   40.41       11   CYS   SG   50.526   49.576   49.063   39.35       12   LYS   N   48.737   51.792   51.812   40.40       12   LYS   CA   47.297   51.914   52.011   40.31       12   LYS   CB   46.926   53.386   52.236   40.50       12   LYS   CG   47.582   54.002   53.467   40.84       12   LYS   CD   47.193   53.242   54.728   41.39       12   LYS   CE   47.795   53.864   55.982   42.05       12   LYS   NZ   47.309   55.259   56.185   42.58       12   LYS   C   46.567   51.353   50.793   40.16       12   LYS   O   46.996   51.573   49.660   39.93       13   LEU   N   45.467   50.637   51.031   39.94       13   LEU   CA   44.698   50.015   49.954   39.87       13   LEU   CB   44.540   48.506   50.189   39.57       13   LEU   CG   45.817   47.718   50.470   39.45       13   LEU   CD1   45.455   46.287   50.855   39.64       13   LEU   CD2   46.742   47.691   49.256   39.49       13   LEU   C   43.283   50.536   49.735   40.21       13   LEU   O   42.590   50.941   50.674   40.15       14   LEU   N   42.861   50.484   48.477   40.14       14   LEU   CA   41.506   50.825   48.098   40.45       14   LEU   CB   41.451   51.674   46.823   40.27       14   LEU   CG   40.026   51.690   46.235   40.37       14   LEU   CD1   39.053   52.388   47.192   40.36       14   LEU   CD2   39.992   52.378   44.869   40.24       14   LEU   C   40.996   49.428   47.775   40.61       14   LEU   O   41.361   48.859   46.749   40.61       15   VAL   N   40.186   48.853   48.652   40.60       15   VAL   CA   39.679   47.518   48.387   40.72       15   VAL   CB   39.685   46.670   49.668   40.68       15   VAL   CG1   39.000   45.333   49.416   40.84       15   VAL   CG2   41.133   46.454   50.110   40.43       15   VAL   C   38.287   47.606   47.805   40.94       15   VAL   O   37.403   48.247   48.374   40.84       16   VAL   N   38.104   46.979   46.648   41.32       16   VAL   CA   36.813   47.000   45.979   41.65       16   VAL   CB   36.908   47.619   44.556   41.38       16   VAL   CG1   35.520   47.653   43.911   41.06       16   VAL   CG2   37.502   49.030   44.633   41.07       16   VAL   C   36.218   45.610   45.844   42.07       16   VAL   O   36.890   44.663   45.430   41.85       17   ALA   N   34.948   45.509   46.217   42.61       17   ALA   CA   34.195   44.272   46.124   43.48       17   ALA   CB   33.525   43.956   47.466   43.31       17   ALA   C   33.140   44.554   45.062   44.03       17   ALA   O   32.409   45.543   45.169   44.04       18   ASP   N   33.073   43.714   44.032   44.77       18   ASP   CA   32.089   43.916   42.973   45.53       18   ASP   CB   32.632   43.438   41.611   45.20       18   ASP   CG   32.781   41.928   41.520   45.15       18   ASP   OD1   32.552   41.243   42.534   45.18       18   ASP   OD2   33.134   41.429   40.426   44.69       18   ASP   C   30.798   43.193   43.329   46.32       18   ASP   O   30.702   42.584   44.398   46.49       19   HIS   N   29.811   43.255   42.440   47.07       19   HIS   CA   28.519   42.626   42.699   47.79       19   HIS   CB   27.505   43.003   41.603   47.47       19   HIS   CG   27.776   42.367   40.273   47.28       19   HIS   CD2   27.273   41.247   39.704   47.04       19   HIS   ND1   28.654   42.902   39.355   47.15       19   HIS   CE1   28.677   42.140   38.275   47.10       19   HIS   NE2   27.848   41.129   38.462   47.20       19   HIS   C   28.585   41.109   42.835   48.24       19   HIS   O   27.810   40.514   43.584   48.59       20   ARG   N   29.496   40.475   42.110   48.97       20   ARG   CA   29.628   39.028   42.194   49.60       20   ARG   CB   30.652   38.517   41.180   50.00       20   ARG   CG   30.268   38.661   39.709   50.36       20   ARG   CD   31.196   37.786   38.863   50.71       20   ARG   NE   30.919   37.837   37.428   50.98       20   ARG   CZ   30.938   38.950   36.705   51.03       20   ARG   NH1   31.217   40.112   37.282   51.22       20   ARG   NH2   30.693   38.900   35.404   51.10       20   ARG   C   30.064   38.606   43.601   49.92       20   ARG   O   29.445   37.743   44.221   50.02       21   PHE   N   31.135   39.223   44.093   50.14       21   PHE   CA   31.680   38.921   45.419   50.40       21   PHE   CB   32.929   39.773   45.671   49.84       21   PHE   CG   33.724   39.353   46.875   49.29       21   PHE   CD1   34.687   38.353   46.773   49.02       21   PHE   CD2   33.512   39.955   48.113   49.00       21   PHE   CE1   35.427   37.961   47.885   48.83       21   PHE   CE2   34.247   39.567   49.227   48.85       21   PHE   CZ   35.206   38.568   49.112   48.76       21   PHE   C   30.645   39.223   46.499   50.80       21   PHE   O   30.398   38.412   47.389   50.79       22   TYR   N   30.057   40.410   46.406   51.47       22   TYR   CA   29.044   40.882   47.344   52.08       22   TYR   CB   28.473   42.208   46.847   52.14       22   TYR   CG   27.346   42.764   47.688   52.33       22   TYR   CD1   27.568   43.176   49.002   52.41       22   TYR   CE1   26.543   43.723   49.768   52.36       22   TYR   CD2   26.061   42.908   47.159   52.31       22   TYR   CE2   25.027   43.451   47.915   52.30       22   TYR   CZ   25.274   43.857   49.219   52.34       22   TYR   OH   24.260   44.396   49.980   52.15       22   TYR   C   27.902   39.890   47.527   52.49       22   TYR   O   27.420   39.678   48.639   52.58       23   ARG   N   27.478   39.285   46.427   52.99       23   ARG   CA   26.377   38.337   46.439   53.57       23   ARG   CB   25.772   38.259   45.032   54.21       23   ARG   CG   24.670   37.231   44.862   55.41       23   ARG   CD   24.194   37.165   43.414   56.15       23   ARG   NE   23.138   36.171   43.243   57.12       23   ARG   CZ   22.521   35.920   42.092   57.61       23   ARG   NH1   22.853   36.592   40.993   57.83       23   ARG   NH2   21.575   34.990   42.037   57.92       23   ARG   C   26.757   36.936   46.915   53.55       23   ARG   O   26.147   36.400   47.841   53.56       24   TYR   N   27.771   36.352   46.288   53.52       24   TYR   CA   28.201   34.997   46.617   53.49       24   TYR   CB   28.807   34.357   45.369   54.15       24   TYR   CG   27.825   34.268   44.218   54.87       24   TYR   CD1   26.763   33.362   44.252   55.28       24   TYR   CE1   25.845   33.285   43.202   55.60       24   TYR   CD2   27.945   35.100   43.103   55.19       24   TYR   CE2   27.031   35.032   42.046   55.54       24   TYR   CZ   25.987   34.122   42.103   55.78       24   TYR   OH   25.096   34.033   41.060   56.16       24   TYR   C   29.156   34.836   47.799   53.15       24   TYR   O   29.227   33.763   48.393   53.19       25   MET   N   29.890   35.889   48.142   52.56       25   MET   CA   30.820   35.816   49.263   51.92       25   MET   CB   32.197   36.350   48.850   51.12       25   MET   CG   32.938   35.473   47.856   50.04       25   MET   SD   33.311   33.822   48.484   48.61       25   MET   CE   34.533   34.186   49.697   49.08       25   MET   C   30.312   36.601   50.468   51.97       25   MET   O   30.580   36.244   51.610   51.92       26   GLY   N   29.583   37.677   50.206   52.06       26   GLY   CA   29.065   38.489   51.289   52.32       26   GLY   C   27.611   38.186   51.579   52.46       26   GLY   O   26.975   38.889   52.360   52.49       27   ARG   N   27.089   37.137   50.949   52.57       27   ARG   CA   25.698   36.740   51.132   52.75       27   ARG   CB   25.506   36.116   52.520   52.63       27   ARG   C   24.799   37.964   50.957   52.81       27   ARG   O   23.786   38.121   51.642   52.84       28   GLY   N   25.192   38.831   50.028   52.75       28   GLY   CA   24.429   40.032   49.748   52.70       28   GLY   C   24.352   41.002   50.906   52.59       28   GLY   O   23.514   41.900   50.912   52.70       29   GLU   N   25.225   40.832   51.890   52.61       29   GLU   CA   25.226   41.716   53.050   52.45       29   GLU   CB   25.178   40.900   54.340   52.72       29   GLU   CG   24.292   39.673   54.284   53.26       29   GLU   CD   24.218   38.966   55.620   53.50       29   GLU   OE1   23.661   39.561   56.564   53.95       29   GLU   OE2   24.721   37.826   55.732   53.73       29   GLU   C   26.462   42.614   53.079   52.17       29   GLU   O   27.596   42.152   52.911   52.18       30   GLU   N   26.237   43.899   53.313   51.57       30   GLU   CA   27.325   44.854   53.365   51.00       30   GLU   CB   26.760   46.264   53.525   51.37       30   GLU   CG   27.780   47.365   53.347   51.94       30   GLU   CD   27.166   48.747   53.451   52.20       30   GLU   OE1   26.101   48.968   52.840   52.34       30   GLU   OE2   27.751   49.614   54.134   52.46       30   GLU   C   28.278   44.539   54.516   50.33       30   GLU   O   29.495   44.505   54.338   50.25       31   SER   N   27.717   44.300   55.697   49.48       31   SER   CA   28.514   44.005   56.880   48.61       31   SER   CB   27.603   43.854   58.105   48.80       31   SER   OG   26.746   42.730   57.967   49.18       31   SER   C   29.353   42.742   56.714   47.87       31   SER   O   30.516   42.710   57.105   47.78       32   THR   N   28.755   41.707   56.134   46.96       32   THR   CA   29.436   40.435   55.924   46.18       32   THR   CB   28.456   39.373   55.407   46.10       32   THR   OG1   27.349   39.281   56.310   46.18       32   THR   CG2   29.132   38.006   55.321   45.75       32   THR   C   30.567   40.599   54.919   45.65       32   THR   O   31.699   40.177   55.165   45.46       33   THR   N   30.254   41.222   53.788   44.84       33   THR   CA   31.248   41.451   52.752   44.08       33   THR   CB   30.637   42.243   51.569   43.91       33   THR   OG1   29.650   41.432   50.922   43.61       33   THR   CG2   31.710   42.623   50.550   43.80       33   THR   C   32.436   42.211   53.333   43.65       33   THR   O   33.585   41.828   53.124   43.57       34   THR   N   32.155   43.271   54.086   43.09       34   THR   CA   33.205   44.091   54.688   42.76       34   THR   CB   32.614   45.317   55.413   42.93       34   THR   OG1   31.956   46.158   54.462   43.33       34   THR   CG2   33.715   46.118   56.100   43.26       34   THR   C   34.082   43.328   55.676   42.38       34   THR   O   35.314   43.433   55.631   42.11       35   ASN   N   33.449   42.566   56.564   41.68       35   ASN   CA   34.183   41.800   57.564   41.10       35   ASN   CB   33.216   41.095   58.521   41.84       35   ASN   CG   32.534   42.065   59.473   42.72       35   ASN   OD1   33.191   42.905   60.091   43.34       35   ASN   ND2   31.212   41.952   59.600   43.12       35   ASN   C   35.126   40.778   56.940   40.39       35   ASN   O   36.269   40.633   57.380   40.26       36   TYR   N   34.647   40.075   55.920   39.29       36   TYR   CA   35.469   39.076   55.255   38.44       36   TYR   CB   34.702   38.447   54.087   38.50       36   TYR   CG   35.481   37.381   53.337   38.59       36   TYR   CD1   36.460   37.729   52.406   38.87       36   TYR   CE1   37.201   36.751   51.737   38.98       36   TYR   CD2   35.258   36.028   53.578   38.76       36   TYR   CE2   35.990   35.042   52.915   38.96       36   TYR   CZ   36.960   35.412   51.999   39.14       36   TYR   OH   37.695   34.448   51.351   39.43       36   TYR   C   36.755   39.728   54.753   37.64       36   TYR   O   37.852   39.220   54.987   37.54       37   LEU   N   36.614   40.871   54.089   36.62       37   LEU   CA   37.764   41.582   53.544   35.66       37   LEU   CB   37.293   42.678   52.589   35.78       37   LEU   CG   36.523   42.090   51.409   35.99       37   LEU   CD1   35.933   43.204   50.540   36.23       37   LEU   CD2   37.423   41.175   50.572   35.84       37   LEU   C   38.692   42.170   54.592   34.90       37   LEU   O   39.908   42.183   54.409   34.78       38   ILE   N   38.125   42.677   55.681   33.72       38   ILE   CA   38.931   43.237   56.750   32.55       38   ILE   CB   38.045   43.881   57.838   32.75       38   ILE   CG2   38.873   44.133   59.101   32.67       38   ILE   CG1   37.404   45.158   57.286   32.95       38   ILE   CD1   36.421   45.792   58.267   33.04       38   ILE   C   39.764   42.117   57.380   32.11       38   ILE   O   40.946   42.304   57.672   31.63       39   GLU   N   39.153   40.951   57.581   31.02       39   GLU   CA   39.888   39.842   58.173   30.48       39   GLU   CB   38.920   38.752   58.652   30.75       39   GLU   CG   38.105   39.226   59.858   30.87       39   GLU   CD   37.639   38.098   60.759   31.45       39   GLU   OE1   38.342   37.065   60.853   31.14       39   GLU   OE2   36.576   38.263   61.402   31.89       39   GLU   C   40.957   39.272   57.236   29.76       39   GLU   O   42.059   38.961   57.681   29.28       40   LEU   N   40.636   39.160   55.948   29.41       40   LEU   CA   41.580   38.648   54.951   29.09       40   LEU   CB   40.910   38.551   53.568   28.82       40   LEU   CG   41.832   38.033   52.437   28.74       40   LEU   CD1   41.002   37.394   51.318   29.05       40   LEU   CD2   42.690   39.160   51.834   28.74       40   LEU   C   42.793   39.581   54.869   29.05       40   LEU   O   43.928   39.124   54.915   29.32       41   ILE   N   42.559   40.886   54.760   28.87       41   ILE   CA   43.666   41.827   54.663   29.03       41   ILE   CB   43.176   43.251   54.326   29.55       41   ILE   CG2   44.336   44.248   54.448   29.61       41   ILE   CG1   42.584   43.247   52.915   29.85       41   ILE   CD1   43.620   42.893   51.871   30.69       41   ILE   C   44.505   41.843   55.920   28.81       41   ILE   O   45.735   41.948   55.843   28.95       42   ASP   N   43.857   41.712   57.082   28.46       42   ASP   CA   44.602   41.692   58.334   28.44       42   ASP   CB   43.642   41.680   59.532   28.61       42   ASP   CG   44.364   41.535   60.858   28.75       42   ASP   OD1   45.085   42.463   61.281   29.10       42   ASP   OD2   44.211   40.477   61.477   29.27       42   ASP   C   45.538   40.474   58.366   28.03       42   ASP   O   46.685   40.580   58.796   27.60       43   ARG   N   45.063   39.324   57.898   28.15       43   ARG   CA   45.906   38.120   57.885   28.17       43   ARG   CB   45.062   36.863   57.623   28.15       43   ARG   CG   44.122   36.454   58.770   27.76       43   ARG   CD   43.472   35.089   58.475   27.71       43   ARG   NE   42.820   35.046   57.161   26.80       43   ARG   CZ   41.529   35.271   56.937   27.38       43   ARG   NH1   40.709   35.562   57.944   27.15       43   ARG   NH2   41.048   35.194   55.696   27.00       43   ARG   C   47.046   38.202   56.857   27.98       43   ARG   O   48.161   37.749   57.125   28.36       44   VAL   N   46.785   38.774   55.690   28.21       44   VAL   CA   47.840   38.909   54.678   28.18       44   VAL   CB   47.269   39.445   53.341   28.21       44   VAL   CG1   48.384   39.630   52.308   27.55       44   VAL   CG2   46.203   38.466   52.821   27.51       44   VAL   C   48.878   39.877   55.238   28.58       44   VAL   O   50.096   39.657   55.139   28.66       45   ASP   N   48.392   40.940   55.865   28.68       45   ASP   CA   49.279   41.922   56.450   28.96       45   ASP   CB   48.460   43.038   57.112   29.15       45   ASP   CG   49.334   44.065   57.813   29.82       45   ASP   OD1   50.246   44.634   57.167   29.51       45   ASP   OD2   49.108   44.298   59.017   29.85       45   ASP   C   50.207   41.239   57.462   29.03       45   ASP   O   51.412   41.502   57.478   28.77       46   ASP   N   49.660   40.348   58.292   29.37       46   ASP   CA   50.483   39.637   59.281   29.71       46   ASP   CB   49.672   38.556   60.019   29.76       46   ASP   CG   48.718   39.133   61.080   30.25       46   ASP   OD1   48.876   40.305   61.490   29.71       46   ASP   OD2   47.818   38.388   61.509   29.87       46   ASP   C   51.693   38.982   58.593   29.78       46   ASP   O   52.816   39.029   59.108   29.62       47   ILE   N   51.460   38.382   57.427   30.23       47   ILE   CA   52.538   37.735   56.679   30.67       47   ILE   CB   51.992   37.002   55.433   31.15       47   ILE   CG2   53.125   36.727   54.447   30.51       47   ILE   CG1   51.279   35.721   55.872   31.48       47   ILE   CD1   50.451   35.132   54.766   32.09       47   ILE   C   53.593   38.755   56.254   31.14       47   ILE   O   54.792   38.510   56.405   30.87       48   TYR   N   53.148   39.896   55.732   31.49       48   TYR   CA   54.068   40.942   55.309   32.49       48   TYR   CB   53.319   42.061   54.583   31.94       48   TYR   CG   53.040   41.766   53.135   31.45       48   TYR   CD1   51.996   40.921   52.760   31.16       48   TYR   CE1   51.788   40.592   51.431   31.44       48   TYR   CD2   53.865   42.281   52.135   31.29       48   TYR   CE2   53.663   41.959   50.801   31.23       48   TYR   CZ   52.629   41.113   50.452   31.44       48   TYR   OH   52.432   40.774   49.131   31.26       48   TYR   C   54.870   41.543   56.460   33.48       48   TYR   O   56.089   41.631   56.384   33.46       49   ARG   N   54.185   41.952   57.526   34.66       49   ARG   CA   54.857   42.550   58.677   36.02       49   ARG   CB   53.841   42.907   59.764   36.26       49   ARG   CG   52.763   43.847   59.289   36.68       49   ARG   CD   53.292   45.229   58.934   37.07       49   ARG   NE   52.209   46.034   58.368   37.56       49   ARG   CZ   52.193   47.364   58.307   37.89       49   ARG   NH1   53.213   48.077   58.781   37.88       49   ARG   NH2   51.140   47.983   57.785   37.74       49   ARG   C   55.942   41.659   59.276   36.73       49   ARG   O   57.010   42.145   59.643   36.86       50   ASN   N   55.674   40.359   59.373   37.87       50   ASN   CA   56.648   39.425   59.938   38.88       50   ASN   CB   55.960   38.130   60.391   39.28       50   ASN   CG   55.127   38.313   61.653   40.06       50   ASN   OD1   55.654   38.613   62.729   40.35       50   ASN   ND2   53.820   38.128   61.527   40.37       50   ASN   C   57.785   39.076   58.981   39.41       50   ASN   O   58.719   38.361   59.355   39.55       51   THR   N   57.716   39.570   57.748   39.83       51   THR   CA   58.759   39.273   56.766   40.21       51   THR   CB   58.193   39.341   55.321   39.71       51   THR   OG1   57.247   38.284   55.130   39.05       51   THR   CG2   59.311   39.214   54.289   39.56       51   THR   C   59.963   40.214   56.884   40.90       51   THR   O   59.811   41.435   56.898   40.72       52   ALA   N   61.160   39.637   56.989   41.77       52   ALA   CA   62.377   40.435   57.081   42.50       52   ALA   CB   63.324   39.846   58.129   42.80       52   ALA   C   63.024   40.422   55.702   43.07       52   ALA   O   63.718   39.478   55.335   43.06       53   TRP   N   62.779   41.481   54.945   43.91       53   TRP   CA   63.289   41.612   53.589   45.05       53   TRP   CB   62.739   42.900   52.974   44.35       53   TRP   CG   61.263   43.028   53.191   44.14       53   TRP   CD2   60.237   42.245   52.570   43.92       53   TRP   CE2   59.009   42.619   53.161   43.92       53   TRP   CE3   60.236   41.260   51.574   43.79       53   TRP   CD1   60.631   43.829   54.104   43.91       53   TRP   NE1   59.280   43.587   54.094   43.79       53   TRP   CZ2   57.790   42.039   52.790   43.71       53   TRP   CZ3   59.020   40.681   51.205   43.76       53   TRP   CH2   57.817   41.076   51.813   43.65       53   TRP   C   64.812   41.568   53.469   46.12       53   TRP   O   65.355   41.475   52.366   46.40       54   ASP   N   65.501   41.633   54.601   47.29       54   ASP   CA   66.957   41.592   54.597   48.59       54   ASP   CB   67.516   42.909   55.136   48.87       54   ASP   CG   67.313   43.057   56.624   49.36       54   ASP   OD1   66.354   42.460   57.157   49.66       54   ASP   OD2   68.106   43.777   57.264   49.88       54   ASP   C   67.405   40.438   55.481   49.24       54   ASP   O   68.549   40.387   55.914   49.44       55   ASN   N   66.486   39.514   55.748   50.07       55   ASN   CA   66.766   38.355   56.590   50.93       55   ASN   CB   67.749   37.415   55.888   51.38       55   ASN   CG   67.176   36.824   54.618   51.78       55   ASN   OD1   66.118   36.188   54.642   52.49       55   ASN   ND2   67.866   37.028   53.500   51.87       55   ASN   C   67.325   38.768   57.948   51.31       55   ASN   O   67.796   37.932   58.719   51.50       56   ALA   N   67.259   40.062   58.238   51.62       56   ALA   CA   67.760   40.590   59.497   51.85       56   ALA   CB   68.875   41.606   59.230   51.88       56   ALA   C   66.641   41.237   60.306   51.99       56   ALA   O   65.756   40.550   60.827   52.12       57   GLY   N   66.683   42.562   60.407   52.02       57   GLY   CA   65.673   43.273   61.172   51.90       57   GLY   C   64.834   44.247   60.371   51.78       57   GLY   O   64.105   45.066   60.941   51.80       58   PHE   N   64.934   44.170   59.047   51.52       58   PHE   CA   64.161   45.056   58.192   51.17       58   PHE   CB   64.863   45.257   56.850   51.44       58   PHE   CG   64.329   46.414   56.053   51.66       58   PHE   CD1   64.497   47.719   56.504   51.71       58   PHE   CD2   63.679   46.201   54.838   51.82       58   PHE   CE1   64.030   48.797   55.754   51.85       58   PHE   CE2   63.206   47.273   54.078   51.76       58   PHE   CZ   63.383   48.573   54.536   51.92       58   PHE   C   62.791   44.427   57.978   50.89       58   PHE   O   62.525   43.813   56.945   50.82       59   LYS   N   61.933   44.565   58.981   50.35       59   LYS   CA   60.584   44.027   58.924   49.76       59   LYS   CB   60.455   42.835   59.875   50.20       59   LYS   CG   60.515   43.213   61.340   50.70       59   LYS   CD   61.263   42.182   62.176   51.68       59   LYS   CE   60.698   40.771   62.023   52.16       59   LYS   NZ   61.171   40.095   60.774   52.55       59   LYS   C   59.623   45.130   59.342   49.05       59   LYS   O   60.017   46.292   59.468   49.13       60   GLY   N   58.366   44.768   59.563   48.23       60   GLY   CA   57.391   45.757   59.973   47.17       60   GLY   C   56.658   46.411   58.821   46.43       60   GLY   O   55.847   47.304   59.041   46.28       61   TYR   N   56.945   45.979   57.594   45.82       61   TYR   CA   56.274   46.529   56.421   45.19       61   TYR   CB   57.231   46.588   55.220   45.23       61   TYR   CG   58.318   47.628   55.362   45.61       61   TYR   CD1   59.495   47.344   56.054   45.69       61   TYR   CE1   60.476   48.310   56.228   46.00       61   TYR   CD2   58.150   48.913   54.844   45.72       61   TYR   CE2   59.128   49.891   55.017   46.07       61   TYR   CZ   60.287   49.579   55.711   46.07       61   TYR   OH   61.254   50.534   55.898   46.61       61   TYR   C   55.030   45.713   56.050   44.60       61   TYR   O   55.018   44.486   56.151   44.29       62   GLY   N   53.982   46.404   55.617   44.24       62   GLY   CA   52.764   45.710   55.245   43.54       62   GLY   C   51.743   46.578   54.547   43.26       62   GLY   O   52.071   47.584   53.916   43.26       63   ILE   N   50.486   46.178   54.674   42.96       63   ILE   CA   49.382   46.884   54.055   42.58       63   ILE   CB   48.708   45.999   52.994   42.40       63   ILE   CG2   49.671   45.795   51.825   42.19       63   ILE   CG1   48.283   44.674   53.628   42.25       63   ILE   CD1   47.610   43.744   52.647   42.44       63   ILE   C   48.350   47.298   55.096   42.70       63   ILE   O   48.451   46.940   56.265   42.36       64   GLN   N   47.353   48.053   54.655   42.99       64   GLN   CA   46.312   48.535   55.549   43.58       64   GLN   CB   46.872   49.646   56.435   43.72       64   GLN   CG   46.027   49.964   57.643   44.56       64   GLN   CD   46.544   51.166   58.404   44.90       64   GLN   OE1   46.220   52.311   58.076   45.06       64   GLN   NE2   47.369   50.915   59.417   45.02       64   GLN   C   45.185   49.080   54.692   43.80       64   GLN   O   45.424   49.864   53.774   43.54       65   ILE   N   43.960   48.654   54.973   44.15       65   ILE   CA   42.821   49.133   54.208   44.77       65   ILE   CB   41.545   48.358   54.549   44.68       65   ILE   CG2   40.369   48.967   53.790   44.65       65   ILE   CG1   41.742   46.876   54.225   44.77       65   ILE   CD1   40.501   46.043   54.484   44.90       65   ILE   C   42.612   50.611   54.517   45.39       65   ILE   O   42.644   51.032   55.673   45.02       66   GLU   N   42.424   51.395   53.466   46.36       66   GLU   CA   42.217   52.825   53.603   47.29       66   GLU   CB   43.081   53.570   52.595   47.91       66   GLU   CG   42.907   55.071   52.625   48.97       66   GLU   CD   44.110   55.764   53.218   49.62       66   GLU   OE1   44.403   55.552   54.422   50.07       66   GLU   OE2   44.774   56.516   52.469   49.95       66   GLU   C   40.757   53.128   53.336   47.78       66   GLU   O   40.147   53.964   54.001   47.77       67   GLN   N   40.211   52.445   52.342   48.29       67   GLN   CA   38.820   52.613   51.971   49.15       67   GLN   CB   38.660   53.773   50.981   49.48       67   GLN   CG   37.210   54.029   50.600   50.14       67   GLN   CD   37.025   55.165   49.609   50.48       67   GLN   OE1   35.895   55.527   49.284   50.88       67   GLN   NE2   38.127   55.729   49.120   50.48       67   GLN   C   38.304   51.330   51.334   49.40       67   GLN   O   39.037   50.640   50.621   49.36       68   ILE   N   37.044   51.010   51.604   49.87       68   ILE   CA   36.419   49.822   51.042   50.50       68   ILE   CB   35.922   48.853   52.135   50.51       68   ILE   CG2   35.176   47.695   51.472   50.47       68   ILE   CG1   37.096   48.365   52.984   50.62       68   ILE   CD1   36.657   47.442   54.122   50.63       68   ILE   C   35.222   50.236   50.203   51.11       68   ILE   O   34.411   51.054   50.625   51.20       69   ARG   N   35.115   49.672   49.010   51.81       69   ARG   CA   34.001   49.973   48.132   52.54       69   ARG   CB   34.488   50.628   46.840   53.05       69   ARG   CG   35.030   52.025   47.035   53.82       69   ARG   CD   34.029   52.901   47.769   54.59       69   ARG   NE   34.453   54.296   47.791   55.44       69   ARG   CZ   34.497   55.078   46.715   55.75       69   ARG   NH1   34.140   54.601   45.528   55.91       69   ARG   NH2   34.897   56.340   46.823   55.87       69   ARG   C   33.268   48.694   47.807   52.75       69   ARG   O   33.845   47.762   47.256   52.71       70   ILE   N   31.992   48.651   48.162   53.11       70   ILE   CA   31.177   47.479   47.898   53.49       70   ILE   CB   30.459   47.005   49.183   53.40       70   ILE   CG2   29.657   45.738   48.885   53.34       70   ILE   CG1   31.484   46.782   50.296   53.46       70   ILE   CD1   30.841   46.428   51.613   53.40       70   ILE   C   30.138   47.815   46.839   53.85       70   ILE   O   29.202   48.566   47.099   53.84       71   LEU   N   30.317   47.271   45.639   54.37       71   LEU   CA   29.379   47.506   44.549   54.89       71   LEU   CB   30.055   47.275   43.198   54.74       71   LEU   CG   31.388   48.015   43.074   54.71       71   LEU   CD1   32.007   47.768   41.699   54.69       71   LEU   CD2   31.210   49.515   43.310   54.57       71   LEU   C   28.229   46.526   44.719   55.34       71   LEU   O   28.326   45.368   44.307   55.27       72   LYS   N   27.144   47.002   45.324   55.90       72   LYS   CA   25.969   46.177   45.587   56.55       72   LYS   CB   24.938   46.986   46.376   56.47       72   LYS   C   25.323   45.575   44.343   56.94       72   LYS   O   24.900   44.421   44.357   56.97       73   SER   N   25.245   46.346   43.265   57.51       73   SER   CA   24.637   45.839   42.038   58.13       73   SER   CB   23.242   46.451   41.846   58.24       73   SER   OG   23.283   47.867   41.883   58.49       73   SER   C   25.497   46.099   40.805   58.43       73   SER   O   26.317   47.018   40.790   58.35       74   PRO   N   25.313   45.287   39.750   58.80       74   PRO   CD   24.286   44.237   39.626   58.89       74   PRO   CA   26.069   45.414   38.501   59.17       74   PRO   CB   25.339   44.466   37.553   59.11       74   PRO   CG   24.798   43.424   38.464   58.95       74   PRO   C   26.069   46.838   37.976   59.58       74   PRO   O   25.188   47.630   38.310   59.54       75   GLN   N   27.067   47.157   37.159   60.04       75   GLN   CA   27.183   48.482   36.565   60.43       75   GLN   CB   28.652   48.859   36.377   60.42       75   GLN   CG   28.870   50.163   35.619   60.40       75   GLN   CD   28.245   51.355   36.315   60.49       75   GLN   OE1   28.527   51.624   37.480   60.43       75   GLN   NE2   27.389   52.080   35.599   60.54       75   GLN   C   26.495   48.464   35.211   60.80       75   GLN   O   26.787   47.614   34.374   60.79       76   GLU   N   25.574   49.397   35.001   61.39       76   GLU   CA   24.859   49.474   33.732   61.84       76   GLU   CB   23.699   50.466   33.841   62.11       76   GLU   CG   22.626   50.078   34.855   62.52       76   GLU   CD   21.468   51.068   34.878   62.88       76   GLU   OE1   20.799   51.224   33.830   63.21       76   GLU   OE2   21.227   51.690   35.937   62.90       76   GLU   C   25.830   49.924   32.643   62.00       76   GLU   O   26.533   50.918   32.811   62.07       77   VAL   N   25.871   49.192   31.534   62.20       77   VAL   CA   26.770   49.531   30.435   62.49       77   VAL   CB   27.862   48.459   30.257   62.52       77   VAL   CG1   28.693   48.369   31.537   62.62       77   VAL   CG2   27.224   47.108   29.924   62.51       77   VAL   C   26.085   49.736   29.083   62.64       77   VAL   O   25.424   48.839   28.544   62.75       78   LYS   N   26.270   50.930   28.535   62.74       78   LYS   CA   25.698   51.291   27.248   62.72       78   LYS   CB   26.135   52.715   26.895   62.87       78   LYS   CG   25.966   53.683   28.068   63.12       78   LYS   CD   26.509   55.069   27.760   63.35       78   LYS   CE   25.682   55.757   26.695   63.52       78   LYS   NZ   26.220   57.097   26.322   63.67       78   LYS   C   26.164   50.296   26.184   62.54       78   LYS   O   27.154   49.584   26.382   62.53       79   PRO   N   25.440   50.216   25.051   62.36       79   PRO   CD   24.173   50.912   24.749   62.31       79   PRO   CA   25.796   49.298   23.964   62.20       79   PRO   CB   24.878   49.745   22.832   62.24       79   PRO   CG   23.626   50.111   23.578   62.28       79   PRO   C   27.273   49.375   23.580   61.91       79   PRO   O   27.788   50.452   23.270   62.02       80   GLY   N   27.947   48.228   23.605   61.55       80   GLY   CA   29.356   48.175   23.258   60.89       80   GLY   C   30.309   48.534   24.385   60.40       80   GLY   O   31.483   48.166   24.331   60.45       81   GLU   N   29.819   49.255   25.391   59.82       81   GLU   CA   30.642   49.675   26.526   59.12       81   GLU   CB   29.969   50.851   27.250   59.16       81   GLU   C   30.926   48.540   27.518   58.67       81   GLU   O   30.174   47.562   27.603   58.69       82   LYS   N   32.021   48.683   28.262   57.81       82   LYS   CA   32.429   47.696   29.263   57.01       82   LYS   CB   33.671   46.933   28.809   57.13       82   LYS   CG   33.473   46.000   27.637   57.26       82   LYS   CD   34.827   45.557   27.082   57.37       82   LYS   CE   35.623   46.743   26.528   57.56       82   LYS   NZ   36.890   46.328   25.854   57.55       82   LYS   C   32.756   48.344   30.597   56.39       82   LYS   O   33.085   49.530   30.678   56.15       83   HIS   N   32.669   47.535   31.643   55.70       83   HIS   CA   32.969   47.970   32.999   55.05       83   HIS   CB   31.767   48.705   33.606   54.78       83   HIS   CG   32.061   49.374   34.916   54.53       83   HIS   CD2   32.328   50.664   35.218   54.59       83   HIS   ND1   32.126   48.682   36.107   54.54       83   HIS   CE1   32.419   49.520   37.085   54.44       83   HIS   NE2   32.548   50.730   36.574   54.55       83   HIS   C   33.291   46.701   33.787   54.71       83   HIS   O   32.661   45.662   33.575   54.68       84   TYR   N   34.276   46.768   34.679   54.17       84   TYR   CA   34.645   45.587   35.455   53.76       84   TYR   CB   35.851   45.880   36.350   52.91       84   TYR   CG   35.579   46.812   37.510   52.25       84   TYR   CD1   35.627   48.199   37.351   51.75       84   TYR   CE1   35.431   49.059   38.438   51.53       84   TYR   CD2   35.316   46.303   38.785   51.78       84   TYR   CE2   35.113   47.150   39.876   51.52       84   TYR   CZ   35.175   48.522   39.701   51.42       84   TYR   OH   35.007   49.343   40.789   50.97       84   TYR   C   33.493   45.059   36.310   53.81       84   TYR   O   33.437   43.867   36.617   53.72       85   ASN   N   32.571   45.941   36.680   54.04       85   ASN   CA   31.437   45.550   37.511   54.28       85   ASN   CB   31.208   46.599   38.604   54.07       85   ASN   CG   30.175   46.160   39.630   53.85       85   ASN   OD1   30.173   45.012   40.072   53.62       85   ASN   ND2   29.302   47.081   40.026   53.78       85   ASN   C   30.133   45.308   36.746   54.72       85   ASN   O   29.061   45.282   37.347   54.66       86   MET   N   30.222   45.121   35.430   55.28       86   MET   CA   29.032   44.865   34.622   55.89       86   MET   CB   29.305   45.136   33.133   56.02       86   MET   CG   30.326   44.225   32.476   56.14       86   MET   SD   30.704   44.761   30.775   56.77       86   MET   CE   30.752   43.196   29.895   56.07       86   MET   C   28.568   43.425   34.813   56.27       86   MET   O   29.360   42.545   35.169   56.28       87   ALA   N   27.283   43.191   34.564   56.60       87   ALA   CA   26.688   41.871   34.735   56.88       87   ALA   CB   25.177   41.953   34.505   56.95       87   ALA   C   27.285   40.784   33.855   57.08       87   ALA   O   27.508   39.662   34.310   57.04       88   LYS   N   27.542   41.107   32.595   57.40       88   LYS   CA   28.097   40.123   31.676   57.94       88   LYS   CB   27.616   40.413   30.251   57.93       88   LYS   C   29.621   40.089   31.715   58.25       88   LYS   O   30.272   41.107   31.956   58.25       89   SER   N   30.187   38.912   31.489   58.63       89   SER   CA   31.633   38.785   31.476   59.25       89   SER   CB   32.059   37.355   31.823   59.17       89   SER   OG   31.725   36.450   30.787   59.39       89   SER   C   32.070   39.146   30.062   59.60       89   SER   O   31.258   39.140   29.136   59.66       90   TYR   N   33.344   39.475   29.900   59.97       90   TYR   CA   33.884   39.847   28.597   60.31       90   TYR   CB   34.038   41.371   28.537   60.49       90   TYR   CG   34.554   41.917   27.225   60.75       90   TYR   CD1   35.901   41.812   26.881   60.73       90   TYR   CE1   36.377   42.321   25.672   60.86       90   TYR   CD2   33.691   42.545   26.326   60.76       90   TYR   CE2   34.156   43.056   25.117   60.81       90   TYR   CZ   35.498   42.942   24.796   60.86       90   TYR   OH   35.957   43.457   23.604   60.88       90   TYR   C   35.238   39.163   28.438   60.43       90   TYR   O   35.975   39.017   29.411   60.46       91   PRO   N   35.597   38.746   27.209   60.56       91   PRO   CD   36.958   38.218   27.000   60.61       91   PRO   CA   34.888   38.845   25.925   60.69       91   PRO   CB   36.017   38.719   24.915   60.67       91   PRO   CG   36.899   37.713   25.577   60.65       91   PRO   C   33.772   37.830   25.645   60.74       91   PRO   O   32.830   38.138   24.908   60.86       92   ASN   N   33.877   36.627   26.208   60.72       92   ASN   CA   32.862   35.594   25.989   60.66       92   ASN   CB   33.482   34.202   26.115   60.91       92   ASN   CG   32.607   33.117   25.509   61.25       92   ASN   OD1   31.443   32.951   25.889   61.45       92   ASN   ND2   33.164   32.372   24.557   61.35       92   ASN   C   31.701   35.713   26.970   60.49       92   ASN   O   31.734   35.126   28.051   60.44       93   GLU   N   30.674   36.463   26.581   60.23       93   GLU   CA   29.502   36.672   27.427   59.90       93   GLU   CB   28.433   37.469   26.667   60.10       93   GLU   C   28.911   35.362   27.919   59.61       93   GLU   O   28.273   35.322   28.974   59.58       94   GLU   N   29.140   34.289   27.166   59.18       94   GLU   CA   28.603   32.976   27.518   58.67       94   GLU   CB   28.631   32.061   26.289   58.81       94   GLU   C   29.302   32.291   28.696   58.28       94   GLU   O   28.686   31.486   29.405   58.33       95   LYS   N   30.579   32.604   28.900   57.63       95   LYS   CA   31.352   32.013   29.986   56.88       95   LYS   CB   32.843   32.047   29.640   56.89       95   LYS   C   31.102   32.719   31.323   56.39       95   LYS   O   30.639   33.863   31.368   56.43       96   ASP   N   31.415   32.026   32.414   55.64       96   ASP   CA   31.230   32.568   33.758   54.84       96   ASP   CB   31.501   31.479   34.805   55.19       96   ASP   CG   31.613   32.036   36.220   55.58       96   ASP   OD1   30.665   32.704   36.685   55.84       96   ASP   OD2   32.650   31.798   36.868   55.61       96   ASP   C   32.100   33.782   34.073   53.98       96   ASP   O   31.639   34.734   34.707   53.84       97   ALA   N   33.348   33.758   33.616   52.96       97   ALA   CA   34.270   34.845   33.916   51.86       97   ALA   CB   35.353   34.325   34.866   52.03       97   ALA   C   34.934   35.558   32.742   51.02       97   ALA   O   35.056   35.020   31.638   50.90       98   TRP   N   35.373   36.784   33.014   49.95       98   TRP   CA   36.080   37.600   32.036   48.91       98   TRP   CB   36.458   38.962   32.608   48.72       98   TRP   CG   35.441   40.023   32.645   48.66       98   TRP   CD2   35.581   41.334   32.094   48.56       98   TRP   CE2   34.458   42.081   32.503   48.57       98   TRP   CE3   36.556   41.953   31.296   48.57       98   TRP   CD1   34.264   40.019   33.335   48.50       98   TRP   NE1   33.671   41.253   33.260   48.39       98   TRP   CZ2   34.279   43.421   32.145   48.50       98   TRP   CZ3   36.379   43.284   30.941   48.55       98   TRP   CH2   35.248   44.004   31.367   48.57       98   TRP   C   37.413   36.939   31.772   48.06       98   TRP   O   37.813   36.005   32.470   47.91       99   ASP   N   38.104   37.453   30.761   47.24       99   ASP   CA   39.457   37.013   30.468   46.10       99   ASP   CB   39.929   37.579   29.137   46.43       99   ASP   CG   41.427   37.508   28.986   46.46       99   ASP   OD1   41.941   36.400   28.727   47.06       99   ASP   OD2   42.092   38.550   29.144   46.60       99   ASP   C   40.144   37.776   31.600   45.23       99   ASP   O   40.003   39.002   31.691   44.88       100   VAL   N   40.867   37.073   32.466   44.29       100   VAL   CA   41.502   37.725   33.605   43.47       100   VAL   CB   42.310   36.706   34.465   43.18       100   VAL   CG1   43.509   36.157   33.689   42.91       100   VAL   CG2   42.758   37.388   35.760   42.85       100   VAL   C   42.387   38.923   33.256   43.23       100   VAL   O   42.304   39.971   33.909   42.97       101   LYS   N   43.225   38.777   32.233   42.98       101   LYS   CA   44.107   39.864   31.822   43.00       101   LYS   CB   45.033   39.393   30.690   43.37       101   LYS   CG   46.058   40.425   30.248   43.86       101   LYS   CD   46.966   39.889   29.141   44.34       101   LYS   CE   47.942   40.968   28.673   44.98       101   LYS   NZ   49.056   40.417   27.830   44.94       101   LYS   C   43.281   41.079   31.377   42.76       101   LYS   O   43.573   42.216   31.754   42.74       102   MET   N   42.245   40.838   30.579   42.66       102   MET   CA   41.385   41.924   30.121   42.38       102   MET   CB   40.387   41.424   29.077   42.94       102   MET   CG   41.000   41.193   27.696   43.54       102   MET   SD   39.812   40.663   26.435   44.26       102   MET   CE   40.520   39.136   25.937   44.13       102   MET   C   40.632   42.531   31.296   41.87       102   MET   O   40.461   43.744   31.373   42.04       103   LEU   N   40.186   41.687   32.220   41.39       103   LEU   CA   39.457   42.177   33.376   40.80       103   LEU   CB   38.980   41.007   34.245   40.97       103   LEU   CG   38.421   41.464   35.605   41.14       103   LEU   CD1   37.354   42.554   35.429   41.16       103   LEU   CD2   37.840   40.278   36.376   40.92       103   LEU   C   40.315   43.124   34.207   40.26       103   LEU   O   39.854   44.181   34.630   39.94       104   LEU   N   41.570   42.751   34.432   39.97       104   LEU   CA   42.457   43.586   35.234   39.64       104   LEU   CB   43.802   42.873   35.453   39.38       104   LEU   CG   44.769   43.686   36.322   39.11       104   LEU   CD1   44.122   44.066   37.656   38.99       104   LEU   CD2   46.064   42.905   36.567   39.19       104   LEU   C   42.673   44.958   34.591   39.70       104   LEU   O   42.686   45.977   35.282   39.74       105   GLU   N   42.829   44.991   33.270   39.72       105   GLU   CA   43.038   46.262   32.590   39.81       105   GLU   CB   43.403   46.048   31.113   39.50       105   GLU   CG   44.044   47.290   30.505   39.45       105   GLU   CD   44.533   47.106   29.080   39.30       105   GLU   OE1   45.020   46.004   28.735   39.29       105   GLU   OE2   44.446   48.082   28.308   39.37       105   GLU   C   41.783   47.130   32.701   40.04       105   GLU   O   41.875   48.339   32.914   39.95       106   GLN   N   40.611   46.510   32.565   40.23       106   GLN   CA   39.351   47.246   32.676   40.32       106   GLN   CB   38.159   46.328   32.436   40.79       106   GLN   CG   36.847   47.083   32.344   41.31       106   GLN   CD   36.810   47.977   31.122   41.76       106   GLN   OE1   36.925   47.497   30.003   42.24       106   GLN   NE2   36.663   49.282   31.331   41.92       106   GLN   C   39.214   47.853   34.066   40.23       106   GLN   O   38.889   49.037   34.225   39.87       107   PHE   N   39.451   47.030   35.080   39.95       107   PHE   CA   39.350   47.502   36.448   39.81       107   PHE   CB   39.753   46.396   37.425   39.46       107   PHE   CG   39.855   46.859   38.843   39.13       107   PHE   CD1   38.759   47.434   39.479   38.92       107   PHE   CD2   41.057   46.745   39.541   39.16       107   PHE   CE1   38.853   47.892   40.783   38.80       107   PHE   CE2   41.166   47.200   40.852   38.87       107   PHE   CZ   40.062   47.777   41.475   39.11       107   PHE   C   40.249   48.714   36.635   39.97       107   PHE   O   39.852   49.705   37.249   39.64       108   SER   N   41.467   48.624   36.100   40.20       108   SER   CA   42.435   49.708   36.198   40.65       108   SER   CB   43.764   49.281   35.555   40.75       108   SER   OG   44.397   48.270   36.326   40.70       108   SER   C   41.901   50.976   35.525   41.00       108   SER   O   42.091   52.085   36.023   40.78       109   PHE   N   41.232   50.796   34.392   41.58       109   PHE   CA   40.664   51.914   33.647   42.31       109   PHE   CB   40.138   51.422   32.294   42.55       109   PHE   CG   39.381   52.468   31.512   43.19       109   PHE   CD1   40.058   53.489   30.840   43.31       109   PHE   CD2   37.986   52.444   31.466   43.13       109   PHE   CE1   39.358   54.472   30.134   43.43       109   PHE   CE2   37.277   53.421   30.766   43.47       109   PHE   CZ   37.965   54.440   30.097   43.28       109   PHE   C   39.530   52.604   34.409   42.61       109   PHE   O   39.489   53.828   34.504   42.49       110   ASP   N   38.616   51.810   34.955   43.11       110   ASP   CA   37.468   52.356   35.672   43.65       110   ASP   CB   36.394   51.278   35.834   43.73       110   ASP   CG   35.826   50.827   34.511   43.66       110   ASP   OD1   35.467   51.703   33.698   44.35       110   ASP   OD2   35.725   49.605   34.280   43.87       110   ASP   C   37.756   52.985   37.026   44.03       110   ASP   O   37.168   54.011   37.367   44.05       111   ILE   N   38.658   52.384   37.796   44.24       111   ILE   CA   38.982   52.903   39.118   44.73       111   ILE   CB   39.388   51.739   40.074   44.96       111   ILE   CG2   40.899   51.509   40.001   45.08       111   ILE   CG1   38.920   52.040   41.499   45.06       111   ILE   CD1   37.407   52.123   41.605   45.37       111   ILE   C   40.097   53.952   39.091   44.84       111   ILE   O   40.495   54.472   40.134   44.79       112   ALA   N   40.581   54.269   37.894   45.28       112   ALA   CA   41.668   55.234   37.703   45.60       112   ALA   CB   41.664   55.721   36.251   45.60       112   ALA   C   41.703   56.437   38.656   46.11       112   ALA   O   42.695   56.654   39.359   45.85       113   GLU   N   40.633   57.230   38.672   46.58       113   GLU   CA   40.579   58.404   39.542   47.25       113   GLU   CB   39.206   59.083   39.453   48.10       113   GLU   CG   38.831   59.503   38.044   49.39       113   GLU   CD   38.447   58.322   37.168   50.14       113   GLU   OE1   38.460   58.476   35.927   50.83       113   GLU   OE2   38.124   57.243   37.721   50.63       113   GLU   C   40.882   58.058   40.996   47.18       113   GLU   O   41.702   58.714   41.640   46.96       114   GLU   N   40.223   57.028   41.515   47.26       114   GLU   CA   40.459   56.624   42.895   47.33       114   GLU   CB   39.387   55.624   43.339   48.04       114   GLU   CG   37.978   56.204   43.279   49.07       114   GLU   CD   36.938   55.295   43.903   49.76       114   GLU   OE1   37.024   55.045   45.129   50.01       114   GLU   OE2   36.034   54.831   43.169   50.18       114   GLU   C   41.859   56.027   43.058   46.96       114   GLU   O   42.580   56.374   43.990   46.83       115   ALA   N   42.250   55.155   42.131   46.60       115   ALA   CA   43.560   54.518   42.188   46.18       115   ALA   CB   43.763   53.636   40.954   46.08       115   ALA   C   44.692   55.540   42.296   45.96       115   ALA   O   45.732   55.261   42.888   45.88       116   SER   N   44.479   56.726   41.734   45.83       116   SER   CA   45.491   57.781   41.752   45.45       116   SER   CB   45.051   58.941   40.857   45.79       116   SER   OG   43.950   59.637   41.431   46.56       116   SER   C   45.783   58.321   43.145   45.01       116   SER   O   46.820   58.942   43.370   44.95       117   LYS   N   44.877   58.078   44.084   44.49       117   LYS   CA   45.034   58.592   45.438   43.96       117   LYS   CB   43.699   59.167   45.916   44.62       117   LYS   CG   43.180   60.294   45.040   45.26       117   LYS   CD   41.749   60.682   45.391   45.71       117   LYS   CE   41.290   61.840   44.512   46.27       117   LYS   NZ   39.905   62.270   44.841   46.64       117   LYS   C   45.562   57.617   46.481   43.42       117   LYS   O   45.813   58.014   47.617   43.44       118   VAL   N   45.735   56.351   46.111   42.52       118   VAL   CA   46.239   55.359   47.062   41.47       118   VAL   CB   45.155   54.330   47.435   41.77       118   VAL   CG1   44.181   54.939   48.444   41.78       118   VAL   CG2   44.417   53.882   46.168   41.66       118   VAL   C   47.457   54.579   46.589   40.74       118   VAL   O   47.823   54.611   45.416   40.46       119   CYS   N   48.076   53.867   47.526   39.88       119   CYS   CA   49.242   53.057   47.226   38.75       119   CYS   C   48.871   51.928   46.269   38.23       119   CYS   O   49.550   51.705   45.262   37.93       119   CYS   CB   49.828   52.489   48.516   38.99       119   CYS   SG   51.156   51.304   48.189   39.08       120   LEU   N   47.782   51.232   46.582   37.29       120   LEU   CA   47.294   50.130   45.758   36.83       120   LEU   CB   47.825   48.787   46.268   36.78       120   LEU   CG   49.289   48.477   45.935   36.84       120   LEU   CD1   49.699   47.143   46.584   36.69       120   LEU   CD2   49.505   48.411   44.416   36.93       120   LEU   C   45.776   50.044   45.758   36.55       120   LEU   O   45.117   50.448   46.712   36.38       121   ALA   N   45.237   49.498   44.676   36.37       121   ALA   CA   43.802   49.283   44.539   36.19       121   ALA   CB   43.233   50.126   43.398   36.11       121   ALA   C   43.670   47.790   44.225   35.94       121   ALA   O   44.370   47.278   43.354   35.76       122   HIS   N   42.789   47.092   44.937   35.85       122   HIS   CA   42.614   45.661   44.717   35.65       122   HIS   CB   43.191   44.869   45.903   35.42       122   HIS   CG   43.523   43.447   45.575   35.09       122   HIS   CD2   42.833   42.302   45.778   34.99       122   HIS   ND1   44.674   43.086   44.906   35.06       122   HIS   CE1   44.675   41.780   44.710   34.72       122   HIS   NE2   43.571   41.280   45.227   34.70       122   HIS   C   41.147   45.298   44.532   35.95       122   HIS   O   40.281   45.718   45.306   35.96       123   LEU   N   40.882   44.503   43.503   36.17       123   LEU   CA   39.536   44.053   43.186   36.35       123   LEU   CB   39.325   44.126   41.676   36.14       123   LEU   CG   38.054   43.424   41.186   36.51       123   LEU   CD1   36.797   44.062   41.781   35.94       123   LEU   CD2   37.995   43.461   39.652   35.81       123   LEU   C   39.250   42.627   43.669   36.77       123   LEU   O   39.970   41.685   43.334   36.61       124   PHE   N   38.204   42.482   44.480   36.97       124   PHE   CA   37.798   41.166   44.955   37.31       124   PHE   CB   37.563   41.158   46.470   36.56       124   PHE   CG   38.827   41.275   47.275   35.87       124   PHE   CD1   39.350   42.520   47.601   35.46       124   PHE   CD2   39.510   40.130   47.683   35.68       124   PHE   CE1   40.536   42.630   48.327   35.35       124   PHE   CE2   40.701   40.225   48.411   35.04       124   PHE   CZ   41.211   41.476   48.732   35.19       124   PHE   C   36.515   40.847   44.222   37.97       124   PHE   O   35.510   41.533   44.395   37.89       125   THR   N   36.576   39.819   43.387   38.53       125   THR   CA   35.442   39.382   42.589   39.62       125   THR   CB   35.743   39.519   41.082   39.80       125   THR   OG1   34.580   39.180   40.313   40.27       125   THR   CG2   36.882   38.572   40.698   40.00       125   THR   C   35.143   37.914   42.872   40.18       125   THR   O   35.818   37.267   43.673   39.87       126   TYR   N   34.129   37.392   42.197   40.95       126   TYR   CA   33.747   35.998   42.363   41.69       126   TYR   CB   32.552   35.879   43.318   41.98       126   TYR   CG   32.259   34.454   43.736   42.49       126   TYR   CD1   33.053   33.808   44.685   42.57       126   TYR   CE1   32.818   32.482   45.039   42.99       126   TYR   CD2   31.218   33.735   43.151   42.74       126   TYR   CE2   30.973   32.403   43.495   42.81       126   TYR   CZ   31.777   31.784   44.437   42.91       126   TYR   OH   31.566   30.465   44.755   43.11       126   TYR   C   33.394   35.384   41.008   42.12       126   TYR   O   32.224   35.230   40.670   42.34       127   GLN   N   34.415   35.065   40.219   42.76       127   GLN   CA   34.208   34.442   38.915   43.26       127   GLN   CB   34.217   35.498   37.799   43.14       127   GLN   CG   35.246   36.612   37.946   42.91       127   GLN   CD   34.958   37.792   37.024   42.59       127   GLN   OE1   34.938   37.654   35.803   42.66       127   GLN   NE2   34.735   38.958   37.610   42.61       127   GLN   C   35.260   33.360   38.672   43.75       127   GLN   O   36.400   33.474   39.120   43.51       128   ASP   N   34.861   32.303   37.973   44.39       128   ASP   CA   35.745   31.179   37.705   45.26       128   ASP   CB   34.920   29.893   37.597   46.00       128   ASP   CG   35.770   28.639   37.682   46.60       128   ASP   OD1   37.011   28.752   37.811   47.14       128   ASP   OD2   35.186   27.533   37.626   47.15       128   ASP   C   36.548   31.386   36.434   45.56       128   ASP   O   36.023   31.228   35.331   45.53       129   PHE   N   37.822   31.740   36.602   45.62       129   PHE   CA   38.719   31.982   35.477   45.80       129   PHE   CB   39.944   32.797   35.916   45.38       129   PHE   CG   39.615   34.179   36.412   44.95       129   PHE   CD1   38.936   35.081   35.599   44.72       129   PHE   CD2   39.986   34.578   37.690   44.59       129   PHE   CE1   38.630   36.362   36.057   44.77       129   PHE   CE2   39.687   35.854   38.158   44.62       129   PHE   CZ   39.006   36.750   37.342   44.61       129   PHE   C   39.193   30.673   34.875   46.19       129   PHE   O   39.149   29.622   35.521   46.15       130   ASP   N   39.651   30.755   33.630   46.61       130   ASP   CA   40.154   29.597   32.896   47.04       130   ASP   CB   40.314   29.949   31.417   47.59       130   ASP   CG   39.089   29.600   30.604   48.18       130   ASP   OD1   38.725   28.403   30.574   48.64       130   ASP   OD2   38.490   30.514   29.994   48.61       130   ASP   C   41.479   29.056   33.412   46.87       130   ASP   O   42.295   29.792   33.967   46.99       131   MET   N   41.682   27.759   33.211   46.72       131   MET   CA   42.907   27.088   33.618   46.61       131   MET   CB   44.112   27.686   32.873   47.71       131   MET   CG   43.962   27.784   31.348   49.21       131   MET   SD   45.510   28.252   30.493   50.81       131   MET   CE   45.787   29.965   31.114   50.14       131   MET   C   43.155   27.148   35.129   45.94       131   MET   O   44.265   26.870   35.582   45.93       132   GLY   N   42.132   27.517   35.899   44.94       132   GLY   CA   42.277   27.580   37.346   44.07       132   GLY   C   43.000   28.794   37.914   43.19       132   GLY   O   43.528   28.745   39.033   43.33       133   THR   N   43.023   29.882   37.150   42.18       133   THR   CA   43.666   31.121   37.578   40.98       133   THR   CB   43.808   32.099   36.383   41.19       133   THR   OG1   44.649   31.515   35.377   41.40       133   THR   CG2   44.414   33.427   36.842   41.23       133   THR   C   42.809   31.771   38.674   39.94       133   THR   O   41.586   31.842   38.546   39.84       134   LEU   N   43.444   32.239   39.747   38.76       134   LEU   CA   42.711   32.869   40.845   37.81       134   LEU   CB   43.042   32.173   42.175   37.94       134   LEU   CG   42.389   30.789   42.318   37.96       134   LEU   CD1   42.685   30.206   43.698   37.70       134   LEU   CD2   40.870   30.873   42.122   37.86       134   LEU   C   42.927   34.377   41.003   37.01       134   LEU   O   42.134   35.065   41.650   36.89       135   GLY   N   43.998   34.902   40.430   36.22       135   GLY   CA   44.235   36.328   40.559   35.22       135   GLY   C   45.271   36.830   39.581   34.68       135   GLY   O   45.902   36.035   38.886   34.19       136   LEU   N   45.449   38.146   39.530   34.01       136   LEU   CA   46.423   38.747   38.625   33.85       136   LEU   CB   45.798   38.886   37.229   33.98       136   LEU   CG   46.809   39.145   36.108   34.21       136   LEU   CD1   47.681   37.906   35.869   34.29       136   LEU   CD2   46.072   39.523   34.814   33.73       136   LEU   C   46.806   40.120   39.182   33.49       136   LEU   O   45.999   40.766   39.852   33.44       137   ALA   N   48.032   40.563   38.912   33.26       137   ALA   CA   48.501   41.858   39.402   33.14       137   ALA   CB   48.891   41.743   40.876   32.82       137   ALA   C   49.691   42.373   38.603   33.33       137   ALA   O   50.378   41.597   37.942   32.89       138   TYR   N   49.932   43.679   38.679   33.35       138   TYR   CA   51.051   44.300   37.967   34.13       138   TYR   CB   50.633   45.642   37.343   33.72       138   TYR   CG   49.420   45.578   36.435   34.20       138   TYR   CD1   49.409   44.750   35.311   34.16       138   TYR   CE1   48.314   44.718   34.444   34.48       138   TYR   CD2   48.295   46.380   36.676   34.16       138   TYR   CE2   47.192   46.359   35.812   34.13       138   TYR   CZ   47.209   45.526   34.700   34.39       138   TYR   OH   46.129   45.474   33.843   34.66       138   TYR   C   52.214   44.554   38.924   34.55       138   TYR   O   52.031   44.643   40.139   34.22       139   GLY   N   53.411   44.688   38.360   35.47       139   GLY   CA   54.582   44.949   39.173   36.62       139   GLY   C   55.701   43.955   38.966   37.31       139   GLY   O   56.866   44.278   39.201   37.52       140   GLY   N   55.364   42.753   38.509   38.05       140   GLY   CA   56.390   41.745   38.301   39.17       140   GLY   C   56.480   41.123   36.915   39.93       140   GLY   O   56.986   40.010   36.772   39.75       141   SER   N   56.005   41.830   35.892   40.71       141   SER   CA   56.047   41.310   34.521   41.34       141   SER   CB   54.726   40.612   34.171   41.61       141   SER   OG   53.644   41.531   34.107   41.68       141   SER   C   56.315   42.424   33.510   41.80       141   SER   O   55.864   43.556   33.690   41.68       142   PRO   N   57.042   42.112   32.418   42.10       142   PRO   CD   57.531   43.131   31.468   42.24       142   PRO   CA   57.606   40.797   32.089   42.58       142   PRO   CB   58.156   41.011   30.681   42.47       142   PRO   CG   58.675   42.413   30.770   42.16       142   PRO   C   58.695   40.313   33.053   42.97       142   PRO   O   59.102   39.148   33.000   43.23       143   ARG   N   59.167   41.204   33.922   43.15       143   ARG   CA   60.210   40.849   34.876   43.34       143   ARG   CB   61.586   41.240   34.315   43.34       143   ARG   C   59.975   41.523   36.230   43.27       143   ARG   O   59.069   42.353   36.380   43.08       144   ALA   N   60.792   41.161   37.217   43.16       144   ALA   CA   60.658   41.746   38.540   43.21       144   ALA   CB   61.685   41.136   39.500   43.24       144   ALA   C   60.846   43.258   38.457   43.31       144   ALA   O   61.550   43.759   37.578   43.14       145   ASN   N   60.197   43.968   39.377   43.45       145   ASN   CA   60.261   45.422   39.472   43.71       145   ASN   CB   61.657   45.857   39.936   43.64       145   ASN   CG   62.076   45.183   41.237   43.64       145   ASN   OD1   62.790   44.185   41.227   43.65       145   ASN   ND2   61.616   45.720   42.359   43.62       145   ASN   C   59.892   46.151   38.183   43.96       145   ASN   O   60.412   47.228   37.904   44.11       146   SER   N   58.986   45.568   37.406   44.30       146   SER   CA   58.561   46.178   36.149   44.53       146   SER   CB   58.053   45.095   35.194   44.90       146   SER   OG   57.994   45.580   33.861   45.57       146   SER   C   57.468   47.231   36.390   44.52       146   SER   O   57.213   47.616   37.536   44.02       147   HIS   N   56.838   47.693   35.306   44.59       147   HIS   CA   55.772   48.698   35.377   44.76       147   HIS   CB   55.110   48.913   34.014   45.09       147   HIS   CG   56.026   49.448   32.966   45.61       147   HIS   CD2   56.441   50.712   32.701   45.76       147   HIS   ND1   56.620   48.642   32.022   45.97       147   HIS   CE1   57.360   49.383   31.216   46.05       147   HIS   NE2   57.266   50.642   31.607   45.97       147   HIS   C   54.667   48.317   36.349   44.59       147   HIS   O   54.346   47.137   36.509   44.78       148   GLY   N   54.072   49.323   36.978   44.20       148   GLY   CA   52.992   49.056   37.905   43.92       148   GLY   C   53.439   48.669   39.297   43.71       148   GLY   O   54.623   48.746   39.643   43.63       149   GLY   N   52.481   48.226   40.101   43.39       149   GLY   CA   52.798   47.862   41.461   43.23       149   GLY   C   52.524   49.052   42.354   43.19       149   GLY   O   51.878   50.010   41.938   42.72       150   VAL   N   53.028   48.994   43.576   43.52       150   VAL   CA   52.813   50.047   44.557   44.13       150   VAL   CB   53.640   49.776   45.836   43.94       150   VAL   CG1   53.225   48.426   46.429   43.80       150   VAL   CG2   55.142   49.794   45.512   43.74       150   VAL   C   53.109   51.464   44.072   44.80       150   VAL   O   53.981   51.685   43.233   44.70       151   CYS   N   52.360   52.416   44.621   45.70       151   CYS   CA   52.490   53.833   44.301   46.52       151   CYS   C   51.926   54.161   42.928   46.62       151   CYS   O   52.421   53.688   41.910   46.64       151   CYS   CB   53.952   54.272   44.383   47.29       151   CYS   SG   54.701   54.094   46.035   48.62       152   PRO   N   50.881   54.996   42.885   46.88       152   PRO   CD   50.302   55.789   43.988   46.91       152   PRO   CA   50.275   55.360   41.608   47.02       152   PRO   CB   49.057   56.178   42.030   47.05       152   PRO   CG   49.553   56.888   43.248   46.98       152   PRO   C   51.227   56.152   40.715   47.22       152   PRO   O   51.923   57.058   41.170   47.34       153   LYS   N   51.271   55.768   39.449   47.40       153   LYS   CA   52.083   56.441   38.445   47.63       153   LYS   CB   53.417   55.730   38.211   48.04       153   LYS   CG   54.161   56.286   36.997   48.80       153   LYS   CD   55.514   55.622   36.766   49.22       153   LYS   CE   56.049   55.963   35.379   49.72       153   LYS   NZ   56.112   57.435   35.118   49.77       153   LYS   C   51.225   56.360   37.199   47.58       153   LYS   O   50.798   55.275   36.804   47.34       154   ALA   N   50.955   57.506   36.588   47.43       154   ALA   CA   50.107   57.533   35.411   47.58       154   ALA   CB   49.631   58.970   35.148   47.52       154   ALA   C   50.751   56.959   34.156   47.66       154   ALA   O   51.945   57.135   33.912   47.36       155   TYR   N   49.935   56.245   33.385   47.93       155   TYR   CA   50.332   55.663   32.108   48.40       155   TYR   CB   50.496   54.145   32.178   48.59       155   TYR   CG   51.659   53.666   33.007   48.89       155   TYR   CD1   51.577   53.627   34.396   49.04       155   TYR   CE1   52.648   53.180   35.172   49.22       155   TYR   CD2   52.846   53.246   32.400   49.00       155   TYR   CE2   53.923   52.797   33.166   49.11       155   TYR   CZ   53.815   52.765   34.553   49.11       155   TYR   OH   54.867   52.313   35.317   48.98       155   TYR   C   49.156   55.981   31.201   48.65       155   TYR   O   48.020   55.591   31.489   48.52       156   TYR   N   49.422   56.695   30.115   48.94       156   TYR   CA   48.362   57.068   29.196   49.32       156   TYR   CB   48.830   58.192   28.261   49.84       156   TYR   CG   47.712   58.733   27.392   50.40       156   TYR   CD1   46.759   59.608   27.915   50.55       156   TYR   CE1   45.697   60.059   27.141   50.83       156   TYR   CD2   47.573   58.322   26.064   50.63       156   TYR   CE2   46.511   58.763   25.283   50.88       156   TYR   CZ   45.579   59.629   25.827   50.96       156   TYR   OH   44.519   60.052   25.059   51.42       156   TYR   C   47.886   55.884   28.368   49.30       156   TYR   O   48.671   55.239   27.673   49.46       157   SER   N   46.595   55.595   28.453   49.12       157   SER   CA   46.010   54.509   27.688   49.10       157   SER   CB   44.894   53.839   28.489   48.89       157   SER   OG   44.305   52.779   27.759   48.59       157   SER   C   45.448   55.103   26.395   49.27       157   SER   O   44.431   55.794   26.412   49.40       158   PRO   N   46.126   54.865   25.259   49.31       158   PRO   CD   47.451   54.230   25.138   49.24       158   PRO   CA   45.680   55.382   23.959   49.35       158   PRO   CB   46.646   54.721   22.986   49.33       158   PRO   CG   47.923   54.721   23.781   49.38       158   PRO   C   44.225   55.043   23.668   49.29       158   PRO   O   43.462   55.901   23.228   49.38       159   VAL   N   43.841   53.796   23.918   49.12       159   VAL   CA   42.464   53.380   23.693   49.15       159   VAL   CB   42.311   51.848   23.780   49.15       159   VAL   CG1   40.836   51.468   23.636   49.23       159   VAL   CG2   43.119   51.183   22.673   49.17       159   VAL   C   41.543   54.018   24.736   49.15       159   VAL   O   40.398   54.357   24.445   49.18       160   GLY   N   42.049   54.178   25.953   49.05       160   GLY   CA   41.242   54.771   27.005   48.98       160   GLY   C   41.118   56.279   26.901   48.70       160   GLY   O   40.251   56.880   27.537   48.86       161   LYS   N   41.979   56.887   26.094   48.39       161   LYS   CA   41.984   58.331   25.914   48.06       161   LYS   CB   40.707   58.782   25.197   48.57       161   LYS   CG   40.574   58.210   23.792   49.22       161   LYS   CD   39.228   58.548   23.165   49.84       161   LYS   CE   39.116   57.933   21.776   50.37       161   LYS   NZ   37.765   58.122   21.153   50.81       161   LYS   C   42.113   59.041   27.252   47.59       161   LYS   O   41.632   60.163   27.417   47.67       162   LYS   N   42.763   58.384   28.210   46.75       162   LYS   CA   42.962   58.973   29.531   46.11       162   LYS   CB   41.665   58.937   30.349   46.11       162   LYS   CG   41.400   57.609   31.058   46.38       162   LYS   CD   40.277   57.753   32.073   46.41       162   LYS   CE   40.057   56.480   32.873   46.71       162   LYS   NZ   38.867   56.602   33.778   46.17       162   LYS   C   44.042   58.230   30.300   45.48       162   LYS   O   44.426   57.128   29.929   45.34       163   ASN   N   44.533   58.839   31.371   44.81       163   ASN   CA   45.544   58.185   32.184   44.41       163   ASN   CB   46.232   59.171   33.120   44.52       163   ASN   CG   47.001   60.220   32.380   44.76       163   ASN   OD1   47.576   59.951   31.328   45.24       163   ASN   ND2   47.032   61.425   32.926   44.78       163   ASN   C   44.912   57.090   33.022   43.96       163   ASN   O   43.740   57.160   33.399   43.97       164   ILE   N   45.702   56.066   33.299   43.24       164   ILE   CA   45.249   54.961   34.114   42.52       164   ILE   CB   44.841   53.761   33.239   42.41       164   ILE   CG2   43.707   54.206   32.318   42.46       164   ILE   CG1   46.040   53.223   32.447   42.23       164   ILE   CD1   46.908   52.260   33.246   41.87       164   ILE   C   46.433   54.627   34.999   42.09       164   ILE   O   47.530   55.157   34.794   41.98       165   TYR   N   46.216   53.777   35.993   41.37       165   TYR   CA   47.300   53.410   36.881   40.63       165   TYR   CB   47.073   54.025   38.268   41.16       165   TYR   CG   46.943   55.538   38.225   41.76       165   TYR   CD1   45.740   56.145   37.852   42.03       165   TYR   CE1   45.632   57.531   37.738   42.27       165   TYR   CD2   48.040   56.361   38.488   41.97       165   TYR   CE2   47.946   57.754   38.373   42.23       165   TYR   CZ   46.735   58.328   37.995   42.58       165   TYR   OH   46.628   59.693   37.847   42.79       165   TYR   C   47.439   51.896   36.943   39.77       165   TYR   O   46.458   51.157   36.790   39.50       166   LEU   N   48.671   51.442   37.143   38.73       166   LEU   CA   48.959   50.015   37.202   37.71       166   LEU   CB   50.128   49.689   36.266   37.45       166   LEU   CG   49.830   50.047   34.805   37.43       166   LEU   CD1   51.057   49.796   33.921   37.38       166   LEU   CD2   48.640   49.243   34.283   37.14       166   LEU   C   49.286   49.560   38.621   37.06       166   LEU   O   49.983   48.566   38.819   36.54       167   ASN   N   48.799   50.312   39.604   36.57       167   ASN   CA   49.013   49.979   41.005   36.02       167   ASN   CB   49.129   51.264   41.835   35.90       167   ASN   CG   47.866   52.122   41.782   35.90       167   ASN   OD1   47.078   52.047   40.835   35.38       167   ASN   ND2   47.682   52.952   42.800   35.49       167   ASN   C   47.783   49.171   41.411   35.89       167   ASN   O   47.042   49.547   42.314   35.95       168   SER   N   47.582   48.042   40.742   35.61       168   SER   CA   46.409   47.231   41.009   35.49       168   SER   CB   45.273   47.715   40.124   35.83       168   SER   OG   45.729   47.807   38.786   36.17       168   SER   C   46.563   45.736   40.813   35.19       168   SER   O   47.544   45.248   40.247   34.75       169   GLY   N   45.548   45.023   41.285   34.71       169   GLY   CA   45.515   43.584   41.179   34.72       169   GLY   C   44.141   43.073   41.582   34.64       169   GLY   O   43.335   43.816   42.145   34.48       170   LEU   N   43.855   41.814   41.281   34.68       170   LEU   CA   42.561   41.251   41.645   34.73       170   LEU   CB   41.607   41.284   40.439   34.66       170   LEU   CG   42.086   40.441   39.249   34.68       170   LEU   CD1   41.551   39.014   39.336   34.63       170   LEU   CD2   41.609   41.063   37.932   35.09       170   LEU   C   42.673   39.833   42.189   34.65       170   LEU   O   43.635   39.103   41.916   34.21       171   THR   N   41.668   39.460   42.969   34.78       171   THR   CA   41.590   38.147   43.579   35.11       171   THR   CB   41.803   38.211   45.115   34.88       171   THR   OG1   43.161   38.556   45.414   34.61       171   THR   CG2   41.444   36.868   45.755   34.57       171   THR   C   40.192   37.606   43.344   35.57       171   THR   O   39.211   38.342   43.444   35.84       172   SER   N   40.101   36.323   43.023   36.15       172   SER   CA   38.806   35.684   42.821   36.77       172   SER   CB   38.679   35.115   41.405   36.76       172   SER   OG   37.490   34.342   41.298   36.64       172   SER   C   38.729   34.548   43.827   37.20       172   SER   O   39.711   33.830   44.031   37.07       173   THR   N   37.578   34.390   44.471   37.77       173   THR   CA   37.430   33.325   45.448   38.35       173   THR   CB   36.824   33.867   46.756   38.23       173   THR   OG1   35.577   34.511   46.480   37.93       173   THR   CG2   37.787   34.877   47.396   37.78       173   THR   C   36.581   32.165   44.915   39.17       173   THR   O   36.047   31.363   45.689   38.97       174   LYS   N   36.463   32.082   43.590   39.81       174   LYS   CA   35.717   31.006   42.948   40.76       174   LYS   CB   34.533   31.558   42.149   40.83       174   LYS   CG   33.657   30.477   41.497   41.18       174   LYS   CD   32.508   31.079   40.709   41.38       174   LYS   CE   31.724   30.009   39.943   41.88       174   LYS   NZ   30.543   30.600   39.218   42.08       174   LYS   C   36.664   30.275   42.006   41.39       174   LYS   O   37.503   30.896   41.359   41.31       175   ASN   N   36.530   28.957   41.932   42.11       175   ASN   CA   37.382   28.158   41.059   42.96       175   ASN   CB   38.770   27.983   41.691   42.77       175   ASN   CG   39.808   27.479   40.698   42.84       175   ASN   OD1   39.721   27.752   39.501   42.69       175   ASN   ND2   40.808   26.759   41.198   42.67       175   ASN   C   36.711   26.807   40.848   43.46       175   ASN   O   36.223   26.195   41.801   43.65       176   TYR   N   36.685   26.353   39.595   44.06       176   TYR   CA   36.055   25.083   39.245   44.41       176   TYR   CB   36.945   23.909   39.676   45.08       176   TYR   CG   38.335   23.920   39.054   45.89       176   TYR   CD1   38.525   23.666   37.688   46.26       176   TYR   CE1   39.810   23.693   37.120   46.51       176   TYR   CD2   39.459   24.199   39.830   46.18       176   TYR   CE2   40.736   24.229   39.270   46.46       176   TYR   CZ   40.903   23.978   37.923   46.52       176   TYR   OH   42.169   24.035   37.392   47.33       176   TYR   C   34.678   25.000   39.914   44.40       176   TYR   O   34.361   24.045   40.626   44.47       177   GLY   N   33.884   26.045   39.704   44.31       177   GLY   CA   32.536   26.102   40.244   43.91       177   GLY   C   32.340   26.213   41.750   43.76       177   GLY   O   31.208   26.389   42.206   43.81       178   LYS   N   33.415   26.122   42.526   43.32       178   LYS   CA   33.296   26.194   43.987   42.94       178   LYS   CB   33.913   24.934   44.617   43.40       178   LYS   CG   33.138   23.658   44.317   44.05       178   LYS   CD   33.875   22.393   44.750   44.34       178   LYS   CE   33.049   21.157   44.389   44.84       178   LYS   NZ   33.794   19.866   44.501   45.03       178   LYS   C   33.940   27.427   44.614   42.19       178   LYS   O   34.745   28.114   43.985   42.28       179   THR   N   33.548   27.719   45.850   41.39       179   THR   CA   34.142   28.824   46.584   40.46       179   THR   CB   33.265   29.251   47.765   40.32       179   THR   OG1   32.033   29.780   47.269   40.47       179   THR   CG2   33.972   30.317   48.605   40.08       179   THR   C   35.431   28.203   47.113   39.85       179   THR   O   35.396   27.107   47.666   39.95       180   ILE   N   36.563   28.877   46.929   38.94       180   ILE   CA   37.839   28.327   47.385   38.10       180   ILE   CB   39.037   29.046   46.706   37.98       180   ILE   CG2   38.852   28.980   45.187   37.87       180   ILE   CG1   39.152   30.489   47.215   37.90       180   ILE   CD1   40.253   31.299   46.509   37.67       180   ILE   C   37.952   28.479   48.893   37.44       180   ILE   O   37.200   29.230   49.495   37.34       181   LEU   N   38.889   27.763   49.496   36.91       181   LEU   CA   39.090   27.836   50.940   36.32       181   LEU   CB   40.127   26.792   51.373   36.29       181   LEU   CG   39.647   25.347   51.160   36.15       181   LEU   CD1   40.729   24.351   51.572   36.24       181   LEU   CD2   38.370   25.075   51.970   36.93       181   LEU   C   39.559   29.236   51.333   35.93       181   LEU   O   40.141   29.951   50.519   35.92       182   THR   N   39.287   29.636   52.572   35.30       182   THR   CA   39.719   30.939   53.054   34.71       182   THR   CB   39.209   31.195   54.488   35.13       182   THR   OG1   37.778   31.307   54.471   35.39       182   THR   CG2   39.823   32.483   55.047   35.65       182   THR   C   41.255   30.998   53.045   34.02       182   THR   O   41.846   32.004   52.650   33.73       183   LYS   N   41.899   29.917   53.472   33.23       183   LYS   CA   43.353   29.876   53.497   32.54       183   LYS   CB   43.843   28.583   54.154   32.61       183   LYS   CG   43.545   27.315   53.371   32.68       183   LYS   CD   44.198   26.114   54.023   32.79       183   LYS   CE   44.147   24.900   53.111   33.20       183   LYS   NZ   44.906   23.737   53.680   33.32       183   LYS   C   43.948   30.004   52.089   32.34       183   LYS   O   45.078   30.476   51.936   31.99       184   GLU   N   43.198   29.574   51.071   31.88       184   GLU   CA   43.667   29.676   49.682   31.79       184   GLU   CB   42.859   28.751   48.753   32.16       184   GLU   CG   42.872   27.283   49.166   32.95       184   GLU   CD   41.977   26.407   48.287   33.41       184   GLU   OE1   40.936   26.893   47.825   33.91       184   GLU   OE2   42.315   25.233   48.072   34.31       184   GLU   C   43.503   31.126   49.239   31.34       184   GLU   O   44.401   31.701   48.610   31.32       185   ALA   N   42.368   31.723   49.596   30.75       185   ALA   CA   42.088   33.121   49.249   30.21       185   ALA   CB   40.708   33.523   49.772   30.06       185   ALA   C   43.172   34.045   49.830   29.95       185   ALA   O   43.603   34.997   49.183   29.57       186   ASP   N   43.598   33.769   51.062   29.71       186   ASP   CA   44.654   34.553   51.695   29.29       186   ASP   CB   44.970   34.007   53.092   29.21       186   ASP   CG   43.869   34.291   54.106   29.44       186   ASP   OD1   42.860   34.927   53.751   28.72       186   ASP   OD2   44.029   33.870   55.268   29.64       186   ASP   C   45.927   34.477   50.856   29.17       186   ASP   O   46.666   35.455   50.721   28.88       187   LEU   N   46.192   33.295   50.316   29.08       187   LEU   CA   47.386   33.079   49.511   29.71       187   LEU   CB   47.648   31.564   49.358   30.11       187   LEU   CG   47.946   30.921   50.728   31.10       187   LEU   CD1   48.287   29.441   50.587   31.30       187   LEU   CD2   49.106   31.640   51.419   31.18       187   LEU   C   47.315   33.783   48.150   29.27       187   LEU   O   48.301   34.350   47.700   29.26       188   VAL   N   46.151   33.768   47.512   29.33       188   VAL   CA   45.991   34.438   46.226   28.96       188   VAL   CB   44.530   34.290   45.694   29.41       188   VAL   CG1   44.379   35.028   44.356   29.10       188   VAL   CG2   44.186   32.805   45.503   29.21       188   VAL   C   46.321   35.929   46.410   28.70       188   VAL   O   47.158   36.507   45.697   28.11       189   THR   N   45.691   36.537   47.404   27.95       189   THR   CA   45.910   37.949   47.670   27.87       189   THR   CB   44.899   38.444   48.725   28.07       189   THR   OG1   43.580   38.244   48.213   27.14       189   THR   CG2   45.083   39.940   48.998   27.79       189   THR   C   47.349   38.302   48.086   27.47       189   THR   O   47.881   39.326   47.650   27.36       190   THR   N   47.971   37.473   48.920   27.06       190   THR   CA   49.349   37.721   49.356   26.95       190   THR   CB   49.855   36.607   50.310   26.97       190   THR   OG1   49.024   36.535   51.472   26.25       190   THR   CG2   51.306   36.879   50.734   26.12       190   THR   C   50.251   37.709   48.114   27.31       190   THR   O   51.143   38.541   47.962   27.08       191   HIS   N   49.988   36.747   47.234   27.47       191   HIS   CA   50.740   36.558   45.991   27.77       191   HIS   CB   50.303   35.248   45.326   27.41       191   HIS   CG   50.934   34.995   43.990   27.50       191   HIS   CD2   50.646   35.485   42.755   27.59       191   HIS   ND1   51.950   34.083   43.810   27.34       191   HIS   CE1   52.259   34.013   42.525   27.71       191   HIS   NE2   51.479   34.853   41.865   27.92       191   HIS   C   50.559   37.717   45.002   27.83       191   HIS   O   51.530   38.199   44.423   28.20       192   GLU   N   49.322   38.147   44.790   27.77       192   GLU   CA   49.086   39.251   43.876   27.96       192   GLU   CB   47.588   39.432   43.627   28.01       192   GLU   CG   46.898   38.207   43.019   28.59       192   GLU   CD   47.664   37.557   41.865   29.60       192   GLU   OE1   48.456   38.259   41.177   29.97       192   GLU   OE2   47.452   36.337   41.634   29.61       192   GLU   C   49.723   40.529   44.439   27.98       192   GLU   O   50.335   41.307   43.687   27.71       193   LEU   N   49.594   40.742   45.751   27.68       193   LEU   CA   50.207   41.908   46.376   27.69       193   LEU   CB   49.813   42.035   47.854   27.93       193   LEU   CG   48.343   42.445   48.037   28.23       193   LEU   CD1   47.999   42.597   49.525   28.74       193   LEU   CD2   48.052   43.752   47.305   28.07       193   LEU   C   51.721   41.788   46.261   27.81       193   LEU   O   52.419   42.799   46.182   27.59       194   GLY   N   52.214   40.548   46.261   27.45       194   GLY   CA   53.640   40.309   46.127   27.67       194   GLY   C   54.145   40.860   44.805   27.89       194   GLY   O   55.214   41.484   44.758   27.74       195   HIS   N   53.387   40.618   43.730   28.34       195   HIS   CA   53.728   41.138   42.409   28.91       195   HIS   CB   52.722   40.684   41.344   28.90       195   HIS   CG   52.885   39.265   40.904   29.19       195   HIS   CD2   51.972   38.284   40.707   29.13       195   HIS   ND1   54.107   38.728   40.555   29.27       195   HIS   CE1   53.940   37.475   40.167   29.41       195   HIS   NE2   52.653   37.181   40.252   29.67       195   HIS   C   53.686   42.669   42.479   29.47       195   HIS   O   54.517   43.357   41.876   29.42       196   ASN   N   52.702   43.197   43.201   29.77       196   ASN   CA   52.554   44.642   43.350   30.47       196   ASN   CB   51.306   44.963   44.180   30.42       196   ASN   CG   50.025   44.913   43.364   30.22       196   ASN   OD1   48.930   44.822   43.919   30.73       196   ASN   ND2   50.154   44.984   42.047   30.10       196   ASN   C   53.784   45.215   44.033   30.97       196   ASN   O   54.210   46.326   43.726   31.07       197   PHE   N   54.332   44.460   44.983   31.16       197   PHE   CA   55.527   44.893   45.683   32.11       197   PHE   CB   55.654   44.211   47.056   32.06       197   PHE   CG   54.897   44.909   48.160   32.49       197   PHE   CD1   53.505   44.971   48.142   32.55       197   PHE   CD2   55.575   45.481   49.233   32.76       197   PHE   CE1   52.797   45.586   49.173   32.63       197   PHE   CE2   54.876   46.102   50.275   32.94       197   PHE   CZ   53.479   46.154   50.243   32.84       197   PHE   C   56.770   44.604   44.839   32.36       197   PHE   O   57.880   44.755   45.317   32.90       198   GLY   N   56.566   44.169   43.597   32.88       198   GLY   CA   57.672   43.913   42.683   33.15       198   GLY   C   58.218   42.507   42.461   33.54       198   GLY   O   59.084   42.313   41.595   33.69       199   ALA   N   57.752   41.522   43.219   33.46       199   ALA   CA   58.275   40.172   43.038   33.97       199   ALA   CB   57.988   39.314   44.278   33.62       199   ALA   C   57.723   39.486   41.802   34.11       199   ALA   O   56.575   39.721   41.399   33.95       200   GLU   N   58.567   38.668   41.173   34.47       200   GLU   CA   58.142   37.884   40.021   34.84       200   GLU   CB   59.177   37.931   38.876   36.00       200   GLU   CG   60.553   37.335   39.156   37.49       200   GLU   CD   61.565   37.624   38.028   38.44       200   GLU   OE1   61.245   37.381   36.848   38.87       200   GLU   OE2   62.680   38.093   38.330   39.55       200   GLU   C   58.024   36.483   40.626   34.62       200   GLU   O   58.230   36.312   41.827   33.71       201   HIS   N   57.687   35.489   39.819   34.34       201   HIS   CA   57.537   34.142   40.346   34.53       201   HIS   CB   56.856   33.266   39.299   33.54       201   HIS   CG   55.433   33.644   39.047   32.67       201   HIS   CD2   54.541   34.303   39.824   32.28       201   HIS   ND1   54.770   33.331   37.881   32.61       201   HIS   CE1   53.530   33.783   37.946   32.40       201   HIS   NE2   53.366   34.377   39.116   32.32       201   HIS   C   58.842   33.498   40.796   35.26       201   HIS   O   59.933   33.867   40.341   35.22       202   ASP   N   58.727   32.559   41.728   36.11       202   ASP   CA   59.894   31.828   42.187   36.98       202   ASP   CB   59.589   31.079   43.488   36.36       202   ASP   CG   59.438   32.010   44.670   35.88       202   ASP   OD1   60.198   32.988   44.737   35.54       202   ASP   OD2   58.575   31.767   45.534   35.71       202   ASP   C   60.210   30.824   41.076   38.08       202   ASP   O   59.331   30.459   40.305   37.52       203   PRO   N   61.473   30.392   40.966   39.66       203   PRO   CD   62.651   30.883   41.699   40.16       203   PRO   CA   61.864   29.418   39.934   40.91       203   PRO   CB   63.353   29.233   40.180   40.99       203   PRO   CG   63.773   30.564   40.727   40.90       203   PRO   C   61.101   28.123   40.180   42.10       203   PRO   O   60.710   27.840   41.313   42.51       204   ASP   N   60.873   27.334   39.141   43.15       204   ASP   CA   60.152   26.079   39.338   44.22       204   ASP   CB   59.428   25.647   38.064   44.53       204   ASP   CG   60.385   25.360   36.925   44.89       204   ASP   OD1   61.006   26.325   36.426   45.32       204   ASP   OD2   60.525   24.179   36.531   45.28       204   ASP   C   61.085   24.958   39.768   44.93       204   ASP   O   62.308   25.133   39.868   44.76       205   GLY   N   60.486   23.798   40.024   45.69       205   GLY   CA   61.263   22.645   40.432   46.42       205   GLY   C   61.906   22.881   41.783   46.99       205   GLY   O   61.328   23.565   42.633   47.23       206   LEU   N   63.097   22.313   41.980   47.22       206   LEU   CA   63.837   22.458   43.235   47.38       206   LEU   CB   64.500   21.129   43.653   47.47       206   LEU   CG   64.965   21.114   45.126   47.66       206   LEU   CD1   63.767   21.238   46.078   47.96       206   LEU   CD2   65.719   19.822   45.451   47.67       206   LEU   C   64.914   23.540   43.111   47.44       206   LEU   O   65.730   23.555   42.179   47.57       207   ALA   N   64.909   24.449   44.070   47.44       207   ALA   CA   65.863   25.536   44.081   47.34       207   ALA   CB   65.447   26.596   43.058   47.44       207   ALA   C   65.849   26.111   45.484   47.38       207   ALA   O   65.042   25.715   46.322   47.46       208   GLU   N   66.755   27.037   45.749   47.42       208   GLU   CA   66.827   27.672   47.056   47.30       208   GLU   CB   68.024   28.621   47.062   48.01       208   GLU   CG   68.202   29.500   48.286   49.08       208   GLU   CD   69.296   30.532   48.053   49.75       208   GLU   OE1   69.087   31.437   47.209   50.27       208   GLU   OE2   70.370   30.430   48.693   50.06       208   GLU   C   65.518   28.437   47.308   46.98       208   GLU   O   65.086   28.599   48.448   46.85       209   CYS   N   64.886   28.885   46.225   46.21       209   CYS   CA   63.641   29.642   46.305   45.87       209   CYS   C   62.385   28.832   46.015   46.10       209   CYS   O   61.297   29.393   45.886   45.97       209   CYS   CB   63.704   30.828   45.344   44.89       209   CYS   SG   64.959   32.048   45.809   43.60       210   ALA   N   62.540   27.517   45.908   46.44       210   ALA   CA   61.421   26.623   45.644   47.08       210   ALA   CB   61.212   26.467   44.142   47.11       210   ALA   C   61.720   25.269   46.277   47.86       210   ALA   O   61.906   24.274   45.581   47.67       211   PRO   N   61.766   25.219   47.620   48.58       211   PRO   CD   61.587   26.356   48.540   48.62       211   PRO   CA   62.048   23.988   48.366   49.20       211   PRO   CB   61.955   24.439   49.823   49.08       211   PRO   CG   62.341   25.897   49.760   48.69       211   PRO   C   61.083   22.849   48.054   50.05       211   PRO   O   60.004   23.048   47.496   49.93       212   ASN   N   61.503   21.642   48.407   51.18       212   ASN   CA   60.706   20.439   48.206   52.30       212   ASN   CB   61.640   19.224   48.139   52.92       212   ASN   CG   62.555   19.117   49.364   53.44       212   ASN   OD1   62.104   18.809   50.471   54.23       212   ASN   ND2   63.844   19.382   49.169   53.84       212   ASN   C   59.773   20.312   49.416   52.71       212   ASN   O   59.964   20.993   50.424   52.73       213   GLU   N   58.783   19.432   49.328   53.24       213   GLU   CA   57.855   19.238   50.442   53.82       213   GLU   CB   56.958   18.033   50.171   54.25       213   GLU   CG   55.856   18.290   49.155   54.86       213   GLU   CD   54.935   17.087   48.983   55.49       213   GLU   OE1   55.417   16.037   48.489   55.75       213   GLU   OE2   53.735   17.182   49.344   55.69       213   GLU   C   58.534   19.065   51.813   53.91       213   GLU   O   58.130   19.704   52.798   54.01       214   ASP   N   59.551   18.202   51.889   53.72       214   ASP   CA   60.255   17.960   53.159   53.41       214   ASP   CB   61.297   16.847   53.000   54.15       214   ASP   CG   62.163   16.644   54.235   54.73       214   ASP   OD1   61.678   16.044   55.225   55.16       214   ASP   OD2   63.341   17.071   54.202   55.21       214   ASP   C   60.936   19.189   53.736   52.79       214   ASP   O   61.118   19.282   54.953   52.99       215   GLN   N   61.335   20.122   52.880   51.88       215   GLN   CA   61.974   21.331   53.383   50.96       215   GLN   CB   62.968   21.910   52.366   51.60       215   GLN   CG   63.931   22.928   52.975   52.22       215   GLN   CD   65.005   22.267   53.828   52.81       215   GLN   OE1   64.703   21.457   54.714   53.24       215   GLN   NE2   66.268   22.605   53.563   52.88       215   GLN   C   60.888   22.363   53.685   49.85       215   GLN   O   61.183   23.472   54.140   49.74       216   GLY   N   59.635   21.989   53.414   48.46       216   GLY   CA   58.515   22.880   53.679   46.62       216   GLY   C   57.625   23.281   52.506   45.32       216   GLY   O   56.666   24.037   52.692   44.87       217   GLY   N   57.928   22.793   51.303   43.79       217   GLY   CA   57.107   23.139   50.151   42.16       217   GLY   C   57.490   24.463   49.506   40.85       217   GLY   O   58.503   25.064   49.856   40.57       218   LYS   N   56.675   24.928   48.566   39.66       218   LYS   CA   56.962   26.180   47.869   38.74       218   LYS   CB   56.289   26.165   46.488   38.96       218   LYS   CG   56.559   24.907   45.658   39.24       218   LYS   CD   58.053   24.587   45.548   39.71       218   LYS   CE   58.283   23.288   44.756   39.75       218   LYS   NZ   59.659   22.715   44.945   40.80       218   LYS   C   56.540   27.456   48.627   37.69       218   LYS   O   55.681   27.412   49.504   37.33       219   TYR   N   57.171   28.579   48.280   36.44       219   TYR   CA   56.861   29.880   48.871   35.08       219   TYR   CB   58.081   30.790   48.826   35.14       219   TYR   CG   59.141   30.373   49.811   35.36       219   TYR   CD1   58.947   30.542   51.184   35.32       219   TYR   CE1   59.897   30.095   52.111   35.38       219   TYR   CD2   60.314   29.749   49.380   35.55       219   TYR   CE2   61.269   29.299   50.294   35.73       219   TYR   CZ   61.055   29.475   51.655   35.85       219   TYR   OH   62.012   29.037   52.552   35.93       219   TYR   C   55.687   30.502   48.117   34.52       219   TYR   O   55.322   30.036   47.035   33.65       220   VAL   N   55.103   31.552   48.689   33.96       220   VAL   CA   53.916   32.183   48.114   33.59       220   VAL   CB   53.419   33.356   49.003   33.34       220   VAL   CG1   54.312   34.585   48.815   32.91       220   VAL   CG2   51.963   33.673   48.659   32.96       220   VAL   C   53.999   32.657   46.665   33.69       220   VAL   O   52.982   32.665   45.961   33.62       221   MET   N   55.189   33.045   46.211   33.66       221   MET   CA   55.338   33.499   44.827   34.39       221   MET   CB   56.464   34.536   44.698   33.60       221   MET   CG   56.167   35.877   45.391   33.08       221   MET   SD   54.466   36.501   45.157   32.05       221   MET   CE   54.422   36.801   43.385   32.38       221   MET   C   55.548   32.371   43.811   35.23       221   MET   O   55.907   32.626   42.659   35.28       222   TYR   N   55.345   31.126   44.232   35.97       222   TYR   CA   55.463   30.014   43.298   36.93       222   TYR   CB   55.432   28.673   44.027   37.59       222   TYR   CG   55.974   27.540   43.187   38.08       222   TYR   CD1   57.327   27.478   42.866   38.44       222   TYR   CE1   57.832   26.463   42.052   38.71       222   TYR   CD2   55.129   26.554   42.678   38.39       222   TYR   CE2   55.621   25.533   41.868   38.73       222   TYR   CZ   56.976   25.498   41.559   39.11       222   TYR   OH   57.471   24.499   40.746   40.00       222   TYR   C   54.219   30.169   42.414   37.37       222   TYR   O   53.140   30.513   42.902   37.08       223   PRO   N   54.354   29.920   41.103   38.04       223   PRO   CD   55.614   29.584   40.418   38.08       223   PRO   CA   53.256   30.042   40.135   38.39       223   PRO   CB   53.979   29.955   38.796   38.48       223   PRO   CG   55.112   29.055   39.095   38.40       223   PRO   C   52.079   29.077   40.200   38.94       223   PRO   O   50.947   29.448   39.890   38.98       224   ILE   N   52.323   27.847   40.618   39.26       224   ILE   CA   51.244   26.886   40.628   40.09       224   ILE   CB   51.400   25.917   39.451   40.48       224   ILE   CG2   50.837   26.567   38.185   40.60       224   ILE   CG1   52.880   25.526   39.317   40.80       224   ILE   CD1   53.147   24.535   38.211   41.41       224   ILE   C   51.056   26.058   41.873   40.24       224   ILE   O   51.989   25.821   42.638   40.30       225   ALA   N   49.816   25.614   42.034   40.58       225   ALA   CA   49.380   24.764   43.127   40.97       225   ALA   CB   49.722   23.308   42.795   41.20       225   ALA   C   49.825   25.070   44.556   41.21       225   ALA   O   49.990   24.139   45.343   41.39       226   VAL   N   50.037   26.330   44.920   41.25       226   VAL   CA   50.394   26.568   46.314   41.45       226   VAL   CB   51.272   27.840   46.493   41.51       226   VAL   CG1   51.068   28.797   45.334   41.64       226   VAL   CG2   50.969   28.515   47.827   41.44       226   VAL   C   49.057   26.630   47.072   41.43       226   VAL   O   48.406   27.669   47.171   41.19       227   SER   N   48.652   25.460   47.564   41.45       227   SER   CA   47.397   25.236   48.289   41.89       227   SER   CB   47.132   23.736   48.353   41.88       227   SER   OG   48.230   23.085   48.982   41.75       227   SER   C   47.298   25.775   49.708   41.96       227   SER   O   46.190   25.967   50.228   42.00       228   GLY   N   48.447   25.990   50.337   41.99       228   GLY   CA   48.466   26.463   51.709   42.37       228   GLY   C   48.642   25.275   52.635   42.72       228   GLY   O   48.721   25.426   53.853   42.58       229   ASP   N   48.710   24.087   52.037   43.09       229   ASP   CA   48.874   22.835   52.772   43.59       229   ASP   CB   48.539   21.636   51.884   44.44       229   ASP   CG   47.071   21.558   51.518   45.07       229   ASP   OD1   46.698   20.559   50.865   46.12       229   ASP   OD2   46.294   22.472   51.863   45.27       229   ASP   C   50.287   22.645   53.291   43.55       229   ASP   O   50.550   21.722   54.060   43.45       230   HIS   N   51.197   23.512   52.865   43.38       230   HIS   CA   52.586   23.411   53.287   43.24       230   HIS   CB   53.455   23.095   52.076   43.74       230   HIS   CG   53.018   21.863   51.355   44.30       230   HIS   CD2   52.116   21.690   50.357   44.53       230   HIS   ND1   53.423   20.601   51.728   44.35       230   HIS   CE1   52.787   19.703   50.997   44.69       230   HIS   NE2   51.988   20.339   50.158   44.94       230   HIS   C   53.064   24.673   53.975   42.89       230   HIS   O   52.631   25.780   53.637   42.41       231   GLU   N   53.963   24.484   54.939   42.49       231   GLU   CA   54.511   25.574   55.735   42.28       231   GLU   CB   55.656   25.064   56.625   43.29       231   GLU   CG   56.270   26.150   57.523   44.54       231   GLU   CD   57.300   25.603   58.521   45.36       231   GLU   OE1   57.909   26.418   59.265   45.54       231   GLU   OE2   57.492   24.365   58.558   45.66       231   GLU   C   55.001   26.786   54.957   41.71       231   GLU   O   54.627   27.915   55.277   41.40       232   ASN   N   55.833   26.563   53.941   40.80       232   ASN   CA   56.381   27.684   53.184   40.30       232   ASN   CB   57.548   27.228   52.289   40.22       232   ASN   CG   58.721   26.673   53.086   40.58       232   ASN   OD1   59.053   27.175   54.157   40.38       232   ASN   ND2   59.364   25.642   52.550   40.48       232   ASN   C   55.359   28.434   52.336   39.58       232   ASN   O   55.616   29.562   51.928   39.26       233   ASN   N   54.215   27.815   52.073   39.33       233   ASN   CA   53.182   28.447   51.252   39.40       233   ASN   CB   51.904   27.605   51.239   39.00       233   ASN   CG   52.081   26.256   50.543   38.74       233   ASN   OD1   51.163   25.441   50.523   38.17       233   ASN   ND2   53.259   26.021   49.973   38.66       233   ASN   C   52.832   29.863   51.702   39.53       233   ASN   O   52.547   30.719   50.871   39.63       234   LYS   N   52.867   30.101   53.012   39.81       234   LYS   CA   52.532   31.403   53.594   40.33       234   LYS   CB   51.901   31.226   54.978   40.90       234   LYS   CG   50.714   30.297   55.069   41.90       234   LYS   CD   50.448   30.006   56.544   42.73       234   LYS   CE   49.282   29.054   56.742   43.39       234   LYS   NZ   49.502   27.769   56.016   44.13       234   LYS   C   53.708   32.361   53.772   40.21       234   LYS   O   53.569   33.363   54.484   40.40       235   MET   N   54.860   32.066   53.173   39.47       235   MET   CA   56.014   32.946   53.333   39.09       235   MET   CB   57.052   32.289   54.244   40.17       235   MET   CG   56.465   31.689   55.512   41.84       235   MET   SD   57.740   30.990   56.579   44.64       235   MET   CE   58.083   29.387   55.761   43.33       235   MET   C   56.674   33.312   52.013   38.33       235   MET   O   56.446   32.657   50.988   37.96       236   PHE   N   57.490   34.360   52.046   37.26       236   PHE   CA   58.204   34.808   50.859   37.15       236   PHE   CB   58.372   36.326   50.881   36.44       236   PHE   CG   57.125   37.077   50.504   35.82       236   PHE   CD1   56.721   37.153   49.172   35.71       236   PHE   CD2   56.344   37.686   51.477   35.44       236   PHE   CE1   55.554   37.832   48.816   35.24       236   PHE   CE2   55.174   38.367   51.132   35.20       236   PHE   CZ   54.777   38.439   49.800   35.07       236   PHE   C   59.577   34.146   50.763   37.28       236   PHE   O   60.243   33.922   51.770   36.90       237   SER   N   59.987   33.823   49.541   37.66       237   SER   CA   61.284   33.205   49.312   38.05       237   SER   CB   61.321   32.542   47.935   38.00       237   SER   OG   61.120   33.505   46.917   37.41       237   SER   C   62.342   34.293   49.357   38.70       237   SER   O   62.024   35.486   49.375   38.35       238   GLN   N   63.601   33.869   49.379   39.50       238   GLN   CA   64.726   34.790   49.380   40.45       238   GLN   CB   66.048   34.010   49.458   41.34       238   GLN   CG   66.243   33.277   50.783   42.67       238   GLN   CD   66.412   34.242   51.951   43.54       238   GLN   OE1   66.160   33.891   53.111   44.24       238   GLN   NE2   66.849   35.468   51.649   43.86       238   GLN   C   64.671   35.590   48.085   40.43       238   GLN   O   65.017   36.769   48.059   40.66       239   CYS   N   64.225   34.939   47.013   40.63       239   CYS   CA   64.120   35.586   45.703   40.74       239   CYS   C   63.102   36.727   45.725   40.41       239   CYS   O   63.356   37.808   45.184   40.04       239   CYS   CB   63.739   34.559   44.628   41.45       239   CYS   SG   65.042   33.335   44.236   42.69       240   SER   N   61.954   36.479   46.349   40.19       240   SER   CA   60.910   37.493   46.462   40.29       240   SER   CB   59.643   36.882   47.071   40.08       240   SER   OG   59.074   35.910   46.215   39.76       240   SER   C   61.383   38.659   47.335   40.40       240   SER   O   61.078   39.823   47.053   40.33       241   LYS   N   62.127   38.339   48.391   40.50       241   LYS   CA   62.637   39.354   49.308   41.10       241   LYS   CB   63.365   38.698   50.481   40.76       241   LYS   CG   62.460   38.012   51.491   40.60       241   LYS   CD   63.282   37.429   52.628   40.39       241   LYS   CE   62.395   36.840   53.707   40.49       241   LYS   NZ   63.204   36.274   54.818   40.45       241   LYS   C   63.586   40.329   48.620   41.77       241   LYS   O   63.494   41.544   48.810   41.57       242   GLN   N   64.505   39.788   47.826   42.60       242   GLN   CA   65.470   40.613   47.113   43.55       242   GLN   CB   66.398   39.712   46.281   44.45       242   GLN   CG   67.611   40.417   45.716   45.86       242   GLN   CD   68.577   39.463   45.038   46.74       242   GLN   OE1   68.199   38.719   44.133   47.71       242   GLN   NE2   69.834   39.486   45.467   47.24       242   GLN   C   64.731   41.607   46.209   43.65       242   GLN   O   65.047   42.797   46.170   43.59       243   SER   N   63.734   41.113   45.485   43.77       243   SER   CA   62.961   41.959   44.588   43.87       243   SER   CB   62.028   41.092   43.740   43.76       243   SER   OG   62.771   40.173   42.961   43.31       243   SER   C   62.145   43.006   45.337   44.23       243   SER   O   62.098   44.171   44.940   44.17       244   ILE   N   61.510   42.588   46.427   44.68       244   ILE   CA   60.669   43.480   47.218   45.21       244   ILE   CB   59.730   42.656   48.137   44.81       244   ILE   CG2   59.051   43.562   49.171   44.62       244   ILE   CG1   58.717   41.909   47.258   44.59       244   ILE   CD1   57.892   40.891   48.014   44.05       244   ILE   C   61.467   44.499   48.025   46.05       244   ILE   O   61.017   45.627   48.224   45.65       245   TYR   N   62.651   44.102   48.481   47.25       245   TYR   CA   63.511   45.006   49.233   48.50       245   TYR   CB   64.825   44.320   49.611   49.13       245   TYR   CG   65.755   45.191   50.434   49.90       245   TYR   CD1   65.463   45.497   51.766   50.03       245   TYR   CE1   66.312   46.302   52.526   50.40       245   TYR   CD2   66.922   45.715   49.878   50.27       245   TYR   CE2   67.781   46.527   50.630   50.62       245   TYR   CZ   67.471   46.815   51.952   50.61       245   TYR   OH   68.321   47.612   52.693   50.69       245   TYR   C   63.799   46.187   48.315   49.10       245   TYR   O   63.675   47.341   48.716   49.04       246   LYS   N   64.174   45.887   47.074   49.81       246   LYS   CA   64.452   46.927   46.090   50.75       246   LYS   CB   64.673   46.323   44.695   51.00       246   LYS   CG   65.982   45.571   44.502   51.59       246   LYS   CD   66.129   45.125   43.052   51.90       246   LYS   CE   67.416   44.357   42.830   52.28       246   LYS   NZ   67.560   43.948   41.403   52.33       246   LYS   C   63.273   47.890   46.023   51.14       246   LYS   O   63.440   49.103   46.118   51.15       247   THR   N   62.078   47.336   45.855   51.80       247   THR   CA   60.866   48.139   45.773   52.40       247   THR   CB   59.614   47.248   45.651   52.30       247   THR   OG1   59.664   46.514   44.420   52.09       247   THR   CG2   58.346   48.107   45.684   52.13       247   THR   C   60.702   49.033   46.998   53.14       247   THR   O   60.542   50.246   46.877   53.08       248   ILE   N   60.750   48.428   48.176   53.94       248   ILE   CA   60.583   49.178   49.408   54.96       248   ILE   CB   60.655   48.240   50.637   54.88       248   ILE   CG2   60.528   49.057   51.925   54.90       248   ILE   CG1   59.562   47.177   50.526   54.89       248   ILE   CD1   59.554   46.217   51.686   54.95       248   ILE   C   61.593   50.314   49.570   55.69       248   ILE   O   61.207   51.454   49.833   55.77       249   GLU   N   62.877   50.027   49.393   56.54       249   GLU   CA   63.878   51.071   49.578   57.48       249   GLU   CB   65.289   50.469   49.652   58.05       249   GLU   CG   65.828   49.905   48.351   59.02       249   GLU   CD   67.257   49.398   48.493   59.54       249   GLU   OE1   68.160   50.222   48.760   59.84       249   GLU   OE2   67.476   48.175   48.345   60.02       249   GLU   C   63.843   52.193   48.543   57.80       249   GLU   O   64.546   53.192   48.693   57.86       250   SER   N   63.025   52.055   47.505   58.16       250   SER   CA   62.953   53.105   46.494   58.45       250   SER   CB   63.514   52.598   45.165   58.61       250   SER   OG   62.729   51.541   44.647   58.94       250   SER   C   61.554   53.672   46.269   58.59       250   SER   O   61.387   54.623   45.506   58.57       251   LYS   N   60.554   53.098   46.935   58.76       251   LYS   CA   59.172   53.554   46.781   58.85       251   LYS   CB   58.351   52.516   46.017   58.99       251   LYS   CG   58.596   52.438   44.520   59.31       251   LYS   CD   57.673   51.377   43.940   59.49       251   LYS   CE   57.609   51.388   42.423   59.66       251   LYS   NZ   56.579   50.404   41.938   59.42       251   LYS   C   58.474   53.839   48.106   58.77       251   LYS   O   57.508   54.597   48.152   58.81       252   ALA   N   58.945   53.217   49.179   58.69       252   ALA   CA   58.339   53.429   50.489   58.69       252   ALA   CB   59.160   52.721   51.573   58.66       252   ALA   C   58.273   54.926   50.772   58.54       252   ALA   O   57.297   55.431   51.331   58.67       253   GLN   N   59.320   55.635   50.364   58.33       253   GLN   CA   59.396   57.071   50.575   57.97       253   GLN   CB   60.815   57.559   50.278   58.55       253   GLN   CG   61.072   59.009   50.627   59.13       253   GLN   CD   60.856   59.299   52.101   59.51       253   GLN   OE1   59.718   59.354   52.577   59.79       253   GLN   NE2   61.952   59.480   52.835   59.76       253   GLN   C   58.399   57.788   49.676   57.38       253   GLN   O   57.889   58.856   50.024   57.51       254   GLU   N   58.114   57.180   48.528   56.50       254   GLU   CA   57.193   57.747   47.551   55.63       254   GLU   CB   57.363   57.039   46.204   56.07       254   GLU   CG   56.591   57.692   45.060   56.47       254   GLU   CD   56.739   56.943   43.746   56.78       254   GLU   OE1   57.891   56.701   43.318   57.11       254   GLU   OE2   55.702   56.602   43.139   56.77       254   GLU   C   55.709   57.735   47.941   54.80       254   GLU   O   55.030   58.753   47.801   54.58       255   CYS   N   55.195   56.602   48.422   53.66       255   CYS   CA   53.778   56.547   48.783   52.59       255   CYS   C   53.410   55.742   50.023   52.52       255   CYS   O   52.231   55.633   50.345   52.30       255   CYS   CB   52.954   56.028   47.603   51.44       255   CYS   SG   53.117   54.239   47.304   49.73       256   PHE   N   54.393   55.171   50.712   52.58       256   PHE   CA   54.095   54.408   51.922   52.90       256   PHE   CB   55.255   53.479   52.288   52.57       256   PHE   CG   55.233   52.163   51.570   52.17       256   PHE   CD1   54.791   52.079   50.252   52.00       256   PHE   CD2   55.701   51.013   52.194   51.99       256   PHE   CE1   54.819   50.872   49.566   51.86       256   PHE   CE2   55.733   49.797   51.512   51.86       256   PHE   CZ   55.291   49.728   50.196   51.82       256   PHE   C   53.832   55.376   53.066   53.32       256   PHE   O   54.473   56.421   53.164   53.19       257   GLN   N   52.888   55.022   53.930   53.81       257   GLN   CA   52.529   55.874   55.057   54.49       257   GLN   CB   51.075   56.327   54.924   54.41       257   GLN   CG   50.729   56.968   53.593   54.72       257   GLN   CD   49.269   57.361   53.509   54.80       257   GLN   OE1   48.826   57.946   52.521   55.05       257   GLN   NE2   48.510   57.038   54.549   54.80       257   GLN   C   52.683   55.125   56.369   54.95       257   GLN   O   53.133   53.981   56.396   55.09       258   GLU   N   52.309   55.788   57.458   55.42       258   GLU   CA   52.353   55.184   58.781   55.77       258   GLU   CB   52.566   56.261   59.851   56.14       258   GLU   CG   53.849   57.070   59.678   56.74       258   GLU   CD   54.055   58.095   60.779   57.19       258   GLU   OE1   55.140   58.729   60.819   57.35       258   GLU   OE2   53.129   58.270   61.606   57.42       258   GLU   C   50.973   54.554   58.949   55.81       258   GLU   O   50.057   54.864   58.185   55.77       259   ARG   N   50.814   53.664   59.921   55.84       259   ARG   CA   49.506   53.058   60.133   55.95       259   ARG   CB   49.577   51.980   61.212   55.87       259   ARG   CG   50.094   50.635   60.726   55.84       259   ARG   CD   50.023   49.614   61.849   55.69       259   ARG   NE   50.405   48.261   61.442   55.63       259   ARG   CZ   49.673   47.461   60.670   55.58       259   ARG   NH1   48.498   47.860   60.196   55.45       259   ARG   NH2   50.113   46.245   60.387   55.58       259   ARG   C   48.504   54.143   60.542   56.13       259   ARG   O   47.427   54.231   59.911   56.19       259   ARG   OXT   48.802   54.904   61.491   56.12       260   007   N3   47.727   29.829   39.980   39.58       260   007   O6   47.576   27.266   41.351   41.05       260   007   O1   44.788   23.897   40.843   43.59       260   007   C5   47.174   31.040   40.173   38.64       260   007   O2   49.740   36.283   39.759   35.36       260   007   C1   46.829   28.736   39.597   40.68       260   007   C2   47.378   28.034   38.342   40.79       260   007   C3   47.601   29.053   37.224   40.76       260   007   C4   48.428   28.495   36.081   40.93       260   007   C6   46.620   27.701   40.709   41.02       260   007   C7   45.036   26.131   41.645   42.36       260   007   C8   44.100   26.443   42.823   42.38       260   007   C13   44.799   27.056   44.045   42.39       260   007   C20   46.068   26.282   44.406   42.54       260   007   C21   43.850   27.067   45.249   42.60       260   007   N1   48.734   35.382   39.482   36.20       260   007   C9   49.235   34.151   39.306   36.48       260   007   O3   50.444   33.937   39.245   35.65       260   007   C10   48.255   33.016   39.175   37.20       260   007   C22   48.177   32.158   40.439   38.16       260   007   C23   47.749   32.989   41.665   37.95       260   007   O4   45.975   31.286   40.126   38.63       260   007   N2   45.330   27.355   40.904   41.60       260   007   C11   44.380   25.075   40.766   42.75       260   007   O5   43.453   25.430   40.003   43.03       260   007   C14   47.884   32.183   42.942   38.16       260   007   C15   49.090   32.237   43.694   38.38       260   007   C16   49.209   31.514   44.917   38.20       260   007   C17   48.120   30.724   45.385   38.22       260   007   C18   46.921   30.645   44.623   38.22       260   007   C19   46.801   31.375   43.401   38.18       260   007   C12   46.392   26.942   37.907   41.12       261   ZN2   ZN + 2   51.599   35.407   39.845   36.08                  
 
         [0155]     The present 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.  
         [0156]     It is further to be understood that all base sizes or amino acid sizes, and all molecular weight or molecular mass values, given for nucleic acids or polypeptides are approximate, and are provided for description.  
         [0157]     Various publications are cited herein, the disclosures of which are hereby incorporated by reference in their entireties.

Technology Category: 8