PATENT DOCUMENT

Abstract:
Soluble polypeptide fraction consisting of all or part one at least of the four immunoglobulin-type extracellular LAG-3 protein domains (amino acids 1-159, 160-230, 240-330 and 331-412 of the SEQ ID NO:1 sequence) or consisting of one peptide sequence derived from these domains by replacement, addition or deletion of one or more amino acids. The fraction of the invention has a specificity at least equal to that of LAG-3 in relation to its ligand.

Full Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a §371 application of PCT/FR95/00593, filed May 5, 1995. 
    
    
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The invention relates to soluble forms derived from the LAG-3 membrane protein which are useful as immunosuppressants, as well as antibodies capable of preventing the specific binding of the LAG-3 protein to MHC (major histocompatibility complex) Class II molecules as immunostimulants. 
     2. Description of the Related Art 
     In WO-A 91/10682, a protein designated LAG-3 has been described. 
     The LAG-3 protein is a protein selectively expressed by NK cells and activated T lymphocytes. Similarity of the amino acid sequence, the comparative exon/intron organization and the chromosomal localization show that LAG-3 is related to CD4. The initial characterization of the LAG-3 gene has been described by TRIEBEL et al. (1). 
     The corresponding DNA codes for a type I transmembrane protein of 498 amino acids containing 4 extra-cellular sequences of the immunoglobulin type. LAG-3 is a member of the immunoglobulin superfamily. 
     The mature protein comprises 476 amino acids (SEQ ID No. 1) with a theoretical molecular weight of 52 kD. The extracellular region contains 8 cysteine residues and 4 potential N-glycosylation sites. By Western blot analysis, it was shown that LAG-3 inside PRA-blasts or activated NK cells has an apparent mass Mr of 70,000. After treatment with N-glycosidase F, a reduction in size to 60 kD was obtained, thereby demonstrating that native LAG-3 is glycosylated. Fuller details are described in WO-A 91/10682. 
     BAIXERAS et al., in J. Exp. Med. 176, 327-337 (2), have, in addition, described their finding that rosette formation between cells transfected with LAG-3 (expressing LAG-3 at their surface) and B lymphocytes expressing MHC Class II was specifically dependent on LAG-3/MHC Class II interaction. 
     Surprisingly, this ligand for MHC Class II was detected with higher levels on activated CD8 +   lymphocytes (MHC Class I-restricted) than on activated CD4 +   lymphocytes. In vivo, only a few disseminated LAG-3 +   cells (MHC Class II-restricted) were to be found in non-hyperplastic lymphoid tissue comprising the primary lymphoid organs, that is to say thymus and bone marrow. LAG-3 +   cells were to be found in hyperplastic lymphoid nodules and tonsils, as well as among peripheral blood mononuclear cells (PBMC) of patients receiving injections of high doses of IL-2. 
     These observations confirm that LAG-3 is an activation antigen in contrast to CD4 expressed in a subpopulation of resting lymphocytes and other cell types, in particular macrophages. 
     The MHC comprises Class I and Class II molecules which are membrane glycoproteins which present fragments of protein antigens to the T lymphocyte receptors (TCR). Class I molecules are responsible for the presentation to CD8 +   cytotoxic cells of peptides derived in large part from endogenously synthesized proteins, while Class II molecules present to CD4 +   helper lymphocytes peptides originating in the first place from foreign proteins which have entered the endocytic, that is to say exogenous, pathway. T helper lymphocytes regulate and amplify the immune response, while cytotoxic lymphocytes are needed to destroy cells irrespective of the tissues expressing &#34;non-self&#34; antigens, for example viral antigens. The mechanism of recognition involves intracellular signals leading to an effective activity of T lymphocytes. 
     It is apparent that, to initiate an immune response mediated by T (CD4 + ) lymphocytes, the foreign antigens must be captured and internalized in the form of peptides by specialized cells, the antigen presenting cells (APC). The resulting antigenic peptides are reexpressed at the surface of the antigen presenting cells, where they are combined with MHC Class II molecules. This MHC Class I/peptide complex is specifically recognized by the T lymphocyte receptor, resulting in an activation of the T helper lymphocytes. 
     Moreover, animal models created by recombination techniques have made it possible to emphasize the part played in vivo by MHC Class II molecules and their ligands. 
     Thus, mice deficient in MHC Class II molecules (3) and possessing almost no peripheral CD4 +   T lymphocytes and having only a few immature CD4 +   lymphocytes in the thymus have proved to be completely incapable of responding to T-dependent antigens. 
     CD4 -  / -   mutant mice (4) have a substantially decreased T lymphocyte activity but show normal development and function of the CD8 +   T lymphocytes, demonstrating that the expression of CD4 on the daughter cells and CD4 +   CD8 +   thymocytes is not obligatory for the development. Compared to normal mice, these CD4-deficient mice have a large amount of CD4 -   CD8 -   cells. 
     These doubly negative cells are restricted to MHC Class II and capable of recognizing the antigen. 
     When they are infected with Leishmania, these mice show a population of functional T helper lymphocytes despite the absence of CD4. These cells are restrictive to MHC Class II and produce interferon-γ when they are activated by the antigen. This indicates that the lineage of the T lymphocytes and their peripheral function need not necessarily depend on the function of CD4. 
     It is now recognized that the proteins encoded by MHC Class II region are involved in many aspects of immune recognition, including the interaction between different lymphoid cells such as lymphocytes and antigen presenting cells. Different observations have also shown that other mechanisms which do not take place via CD4 participate in the effector function of T helper lymphocytes. 
     These different observations underline the pivotal role played by MHC Class II and its ligands in the immune system. 
     Moreover, the importance is known of chimeric molecules composed of the extracytoplasmic domain of proteins capable of binding to ligands and a constant region of human immunoglobulin (Ig) chains for obtaining soluble forms of proteins and of cell receptors which are useful, in particular, as therapeutic agents. 
     Thus, soluble forms of CD4 have proven their efficacy in inhibiting an HIV infection in vitro in a dose-dependent manner. 
     Nevertheless, clinical trials with soluble CD4 molecules, in particular of CD4-Ig, have not enabled a significant decrease in viral titres to be demonstrated. Transgenic mice expressing up to 20 μg/ml of soluble CD4 in their serum were created. These mice showed no difference as regards their immune function relative to control mice. Hitherto, no direct binding to MHC Class II of molecules derived from CD4 has been reported. This strongly suggests that soluble CD4 molecules do not interact in vivo with MHC Class II molecules. 
     SUMMARY OF THE INVENTION 
     Surprisingly, the authors of the present invention have shown that soluble molecules containing different fragments of the extracytoplasmic domain of the LAG-3 protein were capable of binding to MHC Class II molecules and of having an immunosuppressant action. 
     The extracytoplasmic region of LAG-3 represented by the sequence SEQ ID No. 1 comprises the domains D1, D2, D3 and D4 extending from amino acids 1 to 159, 160 to 239, 240 to 330 and 331 to 412, respectively. 
     Thus, the subject of the invention is a soluble polypeptide fraction consisting of all or part of at least one of the 4 immunoglobulin type extracellular domains of the LAG-3 protein (amino acid 1 to 159, 160 to 239, 240 to 330 and 331 to 412 of the sequence SEQ ID No. 1), or of a peptide sequence derived from these domains by replacement, addition and/or deletion of one or more amino acids, and which possesses a specificity at least equal to or greater than that of LAG-3 for its ligand. 
     The present invention encompasses, in particular, soluble polypeptide fractions having a sequence derived from the native LAG-3 sequence originating from the well-known phenomenon of polytypy. 
     The soluble polypeptide fraction is characterized in that it comprises the peptide region of LAG-3 responsible for the affinity of LAG-3 for MHC Class II molecules. 
     The soluble polypeptide fraction comprises, in particular, a peptide sequence derived from these domains by replacement, addition and/or deletion of one or more amino acids, and which possesses a specificity equal to or greater than that of LAG-3 for its ligand, for example the whole of the first two immunoglobulin type domains of LAG-3, or the 4 immunoglobulin type domains of the extracytoplasmic domain of LAG-3. 
     Advantageously, the soluble polypeptide fraction is comprised of all or part of at least one of the four immunoglobulin type extracellular domains of the LAG-3 protein (amino acid 1 to 149, 150 to 239, 240 to 330 and 331 to 412 of sequence SEQ ID No. 1) comprising one or more of the arginine (Arg) rests at the positions 73, 75 and 76 of sequence SEQ ID No. 1 substituted with glutamic acid (Glu). 
     Preferably, the soluble polypeptide fraction comprises a loop in which the average position of the atoms forming the basic linkage arrangement is given by the position of amino acids 46 to 77 (SEQ ID No. 1) appearing in Table 1 or Table 2 or differs therefrom by not more than 5%. 
     The soluble polypeptide fraction advantageously comprises, in addition, the second immunoglobulin type extracellular domain (D2) of LAG-3 (amino acids 150 to 241). 
     Advantageously, the soluble polypeptide fraction comprises, besides the peptide sequence of LAG-3 as defined above, a supplementary peptide sequence at its C-terminal and/or N-terminal end, so as to constitute a fusion protein. The term &#34;fusion protein&#34; means a portion of any protein permitting modification of the physicochemical features of the subfragments of the extracytoplasmic domain of the LAG-3 protein. Examples of such fusion proteins contain fragments of the extracytoplasmic domain of LAG-3 as are defined above, bound to the heavy chain --CH2--CH3 junction region of a human immunoglobulin, preferably an isotype IgG4 immunoglobulin. 
     Such fusion proteins may be dimeric or monomeric. These fusion proteins may be obtained by recombination techniques well known to a person skilled in the art, for example a technique such as that described by Traunecker et al. (5). 
     Generally speaking, the method of production of these fusion proteins comprising an immunoglobulin region fused with a peptide sequence of LAG-3 as defined above consists in inserting into a vector the fragments of cDNA coding for the polypeptide regions corresponding to LAG-3 or derived from LAG-3, where appropriate after amplification by PCR, and the cDNA coding for the relevant region of the immunoglobulin, this cDNA being fused with cDNA coding for the corresponding polypeptide regions or derivatives of LAG-3, and in expressing after transfection the fragments cDNA in an expression system, in particular mammalian cells, for example hamster ovary cells. 
     The fusion proteins according to the invention may also be obtained by cleavage of a LAG-3/ Ig conjugate constructed so as to contain a suitable cleavage site. 
     The subject of the invention is also a therapeutic composition having immunosuppressant activity comprising a soluble polypeptide fraction according to the invention. This composition will be useful for treating pathologies requiring immunosuppression, for example autoimmune diseases. 
     The subject of the invention is also the use of antibodies directed against LAG-3 or soluble polypeptide fractions derived from LAG-3 as are defined above, or fragments of such antibodies, in particular the Fab, Fab&#39; and F(ab&#39;) 2  fragments, for the preparation of a therapeutic composition having immunostimulatory activity. &#34;Immunostimulatory&#34; means a molecular entity capable of stimulating the maturation, differentiation, proliferation and/or function of cells expressing LAG-3, that is to say T lymphocytes or active NK cells. The anti-LAG-3 antibodies may be used as potentiators of vaccines or immunostimulants in immunosuppressed patients, such as patients infected with HIV or treated with immunosuppressant substances, or be used to stimulate the immune system by elimination of self cells displaying abnormal behaviour, for example cancer cells. 
     Immunostimulatory activity of anti-LAG-3 anti-bodies is surprising, inasmuch as anti-CD4 antibodies have an immunosuppressant action. 
     Such antibodies may be polyclonal or monoclonal; however, monoclonal antibodies are preferred. The polyclonal antibodies may be prepared according to well-known methods, such as that described by BENEDICT A.A. et al. (6). Monoclonal antibodies are preferred, on account of the fact that they are specific for a single epitope and yield results with better reproducibility. Methods of production of monoclonal antibodies are well known from the prior art, especially the one described by KOHLER and MILSTEIN. This method, together with variants thereof, are described by YELTON et al. (7). 
     The subject of the invention is also anti-idiotype antibodies directed against the antibodies according to the invention, which contain the internal image of LAG-3 and are consequently capable of binding to MHC Class II. Such antibodies may be used, in particular, as immunosuppressants, and, for example, in autoimmune pathologies. 
     The therapeutic compositions according to the present invention comprise soluble LAG-3 proteins or antibodies as are defined above, as well as a pharmaceutically acceptable vehicle. These compositions may be formulated according to the usual techniques. The vehicle can vary in form in accordance with the chosen administration route: oral, parenteral, sublingual, rectal or nasal. 
     For the compositions for parenteral administration, the vehicle will generally comprise sterile water as well as other possible ingredients promoting the solubility of the composition or its ability to be stored. The parenteral administration routes can consist of intravenous, intramuscular or subcutaneous injections. 
     The therapeutic composition can be of the sustained-release type, in particular for long-term treatments, for example in autoimmune diseases. The dose to be administered depends on the subject to be treated, in particular on the capacity of his/her immune system to achieve the desired degree of protection. The precise amounts of active ingredient to be administered may be readily determined by the practitioner who will initiate the treatment. 
     The therapeutic .compositions according to the invention can comprise, in addition to soluble LAG-3 or the antibodies according to the invention, another active ingredient, where apprto LAG-3 or to anmical bond to LAG-3 or to an antibody according to the invention. As an example, there may be mentioned soluble LAG-3 proteins according to the invention fused to a toxin, for example ricin or diphtheria toxoid, capable of binding to MHC Class II molecules and of killing the target cells, for example leukaemic or melanoma cells, or fused to a radioisotope. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     FIG. 1 shows a comparison of the proliferation of T cells incubated with F(ab) fragments of 17B4 to the proliferation of T cells incubated with intact 17B4 monoclonal antibody. 
     FIG. 2 shows the proliferation of Clone 28 in response to tetanus toxoid when co-cultured with 17B4 or control antibody 10H3. 
     FIG. 3 shows the expression vector pCDM7 used for manufacturing the recombinant LAG-3 proteins and a recombinant CD8 immunoadhesin control. 
     FIG. 4 shows the pCLH3 AXS V2 DHFR hα IVS vector used to express amplified LAG-3 sequences. 
     FIG. 5A shows the inhibition of MHC Class II interaction with LAG-3 by recombinant LAG-3 D1-D4 and FIG. 5B shows the potential inhibition of MHC Class II interaction with CD4. 
     FIG. 6 shows the inhibition of Clone 28 proliferation by recombinant LAG-3 D1-D4. 
     FIG. 7 shows the inhibition of Clone 154 proliferation by LAG-3 Ig. 
     FIG. 8 shows the binding of LAG-3 to B cell lines expressing MHC class II haplotypes or human class II-transfected mouse cells. 
     FIG. 9 shows the binding of LAG-3 Ig to MHC Class II expressing Daudi cells. 
     FIG. 10 shows that preincubation of HLA class II expressing cells with 17B4 inhibits LAG-3 Ig binding. 
     FIGS. 11A-11C show the inhibition of clone T154 proliferation by crosslinked LAG-3 Ig. 
     FIG. 12 compares inhibition of T cell proliferation by anti-Class II antibodies to inhibition of T cell proliferation by LAG-3 Ig. 
     FIGS. 13A-13D show that T cell proliferation in response to OKT3 (FIG. 13A), lectins (FIG. 13B) and low-concentration IL 2  (FIG. 13C) is inhibited by LAG-3 Ig but proliferation in response to high-concentration IL 2  is not inhibited by LAG-3 Ig (FIG. 13D). 
     FIG. 14 shows clone S1B5 cytotoxicity towards Epstein-Barr virus transformed human B cells. 
     FIG. 15 shows peripheral blood lymphocyte cytotoxicity towards HLA Class I -   Daudi cells. 
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     The examples which follow, together with the attached reference figures, will illustrate the invention in greater detail. 
     EXAMPLE 1 
     Proliferation of active T lvmphocvte lines in the presence of anti-LAG-3 monoclonal antibodies 
     The anti-LAG-3 monoclonal antibodies used were 17B4, described in BAIXERAS et al. (2) and deposited at the CNCM under No. I-1240 on Jul. 10, 1992, and 11E3, described in HUARD et al. (8). 
     These antibodies belong to the isotype IgG1. These antibodies were tested for their biological effects on activated T lymphocytes, stimulated by specific antigenic peptides or processed antigens presented by MHC Class II molecules expressed by autologous antigen presenting cells, expressing LAG-3. 
     An anti-CD48 monoclonal antibody designated 10 H3 was used as irrelevant IgG1 antibody (negative control). 
     The saturating concentrations of anti-LAG-3 and anti-CD48 antibodies were determined by immunofluorescence on PHA (phytohaemagglutinin)-blasts and cell lines transformed by Epstein-Barr virus (EBV). In the proliferation tests, the monoclonal antibodies were added in the proportion of 5 times the saturating concentration. 
     The T lymphocyte lines used were, on the one hand the clone 154 derived from peripheral blood lymphocytes, raised against a peptide mimicking an influenza haemagglutinin (HA) fragment having an amino acid sequence extending from amino acid 306 to 329 (p20 peptide), and on the other hand the clone 28, a T lymphocyte clone derived from peripheral lymphocytes of a single human donor, raised against diphtheria toxoid (DT). The antigen presenting cells (APC) corresponding to clone 154 were EBV-transformed B lymphocytes of the same donor (DR3/DR11) as T 154. The antigen presenting cells corresponding to clone 28 were EBV-transformed B lymphocytes of the same donor. This clone was restricted to HLA DR7. 
     For clone 154, the APC (5×10 6 ) were incubated at 37° C. for one and a half hours with variable doses of the p20 peptide, then washed and irradiated (10,000 rad). The cells were plated out on 96-well microtitration plates at the same time as the clone 154 cells (0.5×10 5  to 10×10 5  cells/ml) in a 3:1 ratio. For clone 28, the responding cells/stimulating cells ratio was 1. 
     The HLA DR7/EBV APC cells were either treated with mitomycin or irradiated, then added to the T lymphocytes in the presence of DT (which remained in the culture). The final concentration of clone 28 cells was 100,000 cells/ml. 
       3  H!Thymidine (1 μCi/well) was added at varying time intervals from day 2 to day 10 of culture. 
     Each experiment was carried out in triplicate. 
     The results were expressed as the mean cpm and after subtraction of the cpm found in the negative control (T lymphocytes cocultured with APC unladen with immunogens). The proliferation tests were carried out on 96-well plates. The absorption of tritiated thymidine in the individual 200 μl wells was measured after adding 1 μCi of thymidine for the last 18 hours of culture. The results were expressed in the form of the mean of 3 tests. The standard deviation was usually less than 12% (a little more in the case of very low cpm measurements). Moreover, mixed culture (clone 154/APC) supernatants were combined, filtered through 0.22 μm membranes, divided into samples and frozen at -20° C. until the time of titration using commercial immunoassay kits: Immunotech IL-2 and INF-α titration kit, Genzyme IFN-γ kit and Cayman Chemicals IL-4 kit. 
     A dose determination study was carried out to establish the proliferation profiles of clone 154 brought into contact with the p20 specific antigen at varying concentrations and in the presence or absence of anti-LAG-3 monoclonal antibodies or irrelevant monoclonal antibodies (negative control). 
     The individual results of 16 separate tests showed that, irrespective of the concentration of added antigen, the initial point up to the peak of proliferation was not modified, but a significant prolongation of the proliferation of T lymphocytes incubated with the anti-LAG-3 monoclonal antibodies was observed systematically. Fab fragments of the monoclonal antibody 17B4 were prepared and used in a test of proliferation of clone 154. The proliferation profile of T lymphocytes activated by the antigen with the 17B4 Fab fragments (15 μg/ml) was similar to that of cells incubated in the presence of whole 17B4 monoclonal antibody (40 μg/ml) (FIG. 1). These results show that the observed biological effects are not attributable to a non-specific reaction induced by the Fc region of the anti-LAG-3 monoclonal antibodies. 
     Similar results were obtained with the 11E3 anti-LAG-3 monoclonal antibodies. 
     Clone 28 was also stimulated with the antigen (tetanus toxoid 10 μg/ml) in the presence of 17B4 monoclonal antibodies after coculture with the corresponding APC in the presence of DT. The results are shown in FIG. 2. 
     The effects of the anti-LAG-3 monoclonal antibodies observed with clone 28, namely the prolongation of proliferation, are similar to those observed with clone 154. 
     Tests were carried out designed to measure the miscellaneous cellular events occurring after the antigenic stimulation of clone 154 cells incubated in the presence of anti-LAG-3 monoclonal antibodies. 
     The cells were harvested during conventional antigenic stimulation of clone 154 in the presence of anti-LAG-3 or anti-CD48 monoclonal antibodies or in the absence of antibodies, and tested for the expression of LAG-3 and CD25 transmembrane receptors, and samples of culture supernatants were collected at different time intervals after stimulation and tested for the presence of IFN-γ, TNF-α, IL-4 and IL-2. 
     Two-colour direct immunofluorescence tests (anti-CD3 monoclonal antibodies and anti-CD25 monoclonal antibodies) showed that IL-2 receptors were weakly but significantly increased 5 days after the antigenic stimulation. Similar tests with anti-CD3 and 11E3 (anti-LAG-3) monoclonal antibodies showed that LAG-3 was over-expressed from the day following activation onwards. In addition, the secretion of IL-2, IL-4, IFN-γ and TNF-α was also modulated by incubation with anti-LAG-3 monoclonal antibodies, thus showing that different cellular events are modified by the presence of anti-LAG-3 monoclonal antibodies and that some events already take place 24 hours after stimulation. 
     These results show indirectly that LAG-3 plays a regulatory role for CD4 +   cells. The fact that anti-LAG-3 monoclonal antibodies increase proliferation, and hence act as immunopotentiators, suggest that LAG-3 is involved in the &#34;deactivation&#34; of CD4 +   T lymphocytes with a negative role of LAG-3 on the antigen-dependent stimulation. 
     EXAMPLE 2 
     Transient expression of LAG-3 fusion proteins 
     Soluble proteins derived from LAG-3 were obtained by a recombinant DNA technique using suitable vectors comprising DNA coding for LAG-3 and DNA coding for an immunoglobulin fragment. The transient expression system consisted of transfected Cos cells. This system makes it possible to produce several mg of recombinant fusion proteins. Recombinant DNA techniques were carried out as described by MANIATIS et al. (22). The modifications were made as recommended by the manufacturer. 
     Construction of LAG-3 D1-D4 Ig and LAG-3 D1D2 Ig 
     Fragments coding for the D1D2 or D1-D4 regions were amplified (30 cycles) from a fragment of cDNA (FDC sequence) encompassing LAG-3 cDNA (TRIEBEL et al. (1)), using Taq polymerase free from 5&#39;-endonuclease activity and relatively resistant to an exposure to very high temperature; the amplification was followed by a denaturation at 98° C. (with a Perkin Elmer Cetus &#34;DNA thermal cycle&#34;). Specific primers were used as recorded in the table below. 
     The resulting amplified fragments (739 bp and 1312 bp for LAG-3 D1-D2 and LAG-3 D1-D4, respectively) were inserted into a pBS plasmid (Stratagene). 
     Inserts were prepared after digestion with XhoI and BglII and introduced into the XhoI/BamHI sites of the vector pCDM7-CD8-IgG1 (pCDM7 being derived from pCDM8 marketed by Stratagene), as illustrated in FIG. 3, so as to exchange the DNA sequences coding for CD8 for those coding for the subfragments of LAG-3. The resulting expression vectors contained the sequences coding for D1D2 or D1-D4 fused to the DNA sequences coding for the --CH 2  --CH 3  junction region of a human IgG1 chain. 
     
                                           TABLE 3__________________________________________________________________________Primers used to amplify LAG-3 DNA sequences by PCR                                Resulting                                encoded subfrag-                                ment fused withPrimers used for amplification of the DNA                                a subfragment Ig__________________________________________________________________________Primer (5&#39;)                          LAG-3 D1D25&#39; GCGCCTCGAGGCCCAGACCATAGGAGAGATGT 3&#39; (SEQ ID NO: 2)                                from the leadercouplinguntranslated      start of                  sequence tosite 5&#39; sequences      translation               amino acid 241Primer (3&#39;)5&#39; GCGCAGATCTCTCCAGACCCAGAACAGTGAGGTTATACAT 3&#39; (SEQ ID NO: 3)BglII coup-  End of D2ling sitePrimer (5&#39;)                          LAG-3 D1-D4identical to LAG-3 D1D2              from the leaderPrimer (3&#39;)                          sequence to5&#39; GCGCAGATCTACCTGGGCTAGACAGCTCTGTGAA 3&#39; (SEQ ID NO: 4)                                amino acid 412Bg1II coup-  End of D4ling site__________________________________________________________________________ 
    
     CDM7 is a eukaryotic expression vector derived from the vectors developed by SEED et al. (10) for the cloning of DNA and its expression in E. coli and eukaryotic cells. CDM7 possesses the following features: (i) the human cytomegalovirus promoter for transient expression in mammalian cells; (ii) a viral origin of SV40 for an autosomal replication of mammalian cells expressing T antigen; (iii) πVX (type Col E1) as plasmid origin for a high copy number; (iv) a Sup F selection for resistance to ampicillin and tetracycline in Tet amb  and Amp amb  E. coli strains; (v) an origin of replication of M13 for the release of a single strand; (vi) a T7 RNA promoter; and (vii) a polylinker for an efficient cloning of heterologous DNA. 
     Transient expression in Cos cells 
     Cos cells (5×10 6 ) were transfected with 30 μg of DNA of suitable expression vectors (coding for either LAG-3 D1D2 Ig, or LAG-3 D1-D4 Ig, or CD8 Ig) by electroporation (200 V, 1500 μF, 30-40 msec) using a Cellject apparatus (Eurogentech, Liege, BE). The cells were plated out again and cultured on a medium containing 5% of foetal calf serum. The supernatants were withdrawn 6 days after transfection. 
     The synthesis of the resulting fusion proteins was analysed from the supernatants as well as from cell extracts of transfected cells, by Western blot analysis with the 17B4 monoclonal antibodies. Immunoreactive materials were observed in the supernatant of cells transfected with DNA coding for LAG-3 D1D2 Ig or LAG-3 D1-D4 Ig. 
     Concomitantly, a recombinant CD8 immunoadhesin (CD8 Ig) was obtained as negative control using the same expression system and the expression vector pCDM7-CD8 (FIG. 3). 
     The recombinant proteins LAG-3 D1D2 Ig, LAG-3 D1-D4 Ig and CD8 Ig were purified by means of the standard method on protein A-Sepharose. The resulting material was analysed by SDS-PAGE, followed by Coomassie staining or a Western blot analysis using anti-human Ig antibody. 
     EXAMPLE 3 
     Production of soluble subfragments of LAG-3 
     In order to produce large amounts of recombinant proteins, a stable expression system consisting of transfected mammalian cells was developed. The host cells are anchorage-dependent hamster ovary (CHO) cells isolated from CHO cells deficient in dihydrofolate reductase (DHFR) and consequently necessitating glycine, a purine and thymidine for their growth. The pivotal role of DHFR in the synthesis of nucleic acid precursors, combined with the sensitivity of DHFR-deficient cells with respect to tetrahydrofolate analogues such as methotrexate (MTX), has two major advantages. Transfection of these cells with expression vectors containing the DHFR gene permits the secretion of recombinant DHFR-resistant clones, and the culturing of these cells on selective media containing increasing amounts of MTX results in amplification of the DHFR gene and the DNA associated therewith. 
     Construction of LAG-3 D1, LAG-3 D1D2, LAG-3 D1-D4 
     Fragments of DNA coding for the D1, D1D2 or D1-D4 regions were amplified using a PCR method identical to the one described previously, using the primers specified in the table below. 
     
                                           TABLE 4__________________________________________________________________________Primers used for amplifying LAG-3 DNA sequences by PCR                                Resulting                                encodedPrimers used for amplification of the DNA                                subfragment__________________________________________________________________________Primer (5&#39;)                          LAG-3 D15&#39; CGCCGTCGACCGCTGCCCAGACCATAGGAGAGATGTG 3&#39; (SEQ ID NO:                                from the leaderSalI coup-untranslated      start of                  sequence toling site5&#39; sequences      translation               amino acid 149Primer (3&#39;)5&#39; GCGCGTCGACTTAACCCAGAACAGTGAGGTTATAC 3&#39; (SEQ ID NO: 6)SalI coup-  End of D1ling sitePrimer (5&#39;)                          LAG-3 D1D2identical to LAG-3 D1                from the leaderPrimer (3&#39;)                          sequence to5&#39; GCGCGTCGACTTAACCCAGAACAGTGAGGTTATAC 3&#39; (SEQ ID NO: 7)                                amino acid 239SalIII coup-  End of D2ling sitePrimer (3&#39;)                          amino acid 1495&#39; GCGCGTCGACTTAACCCAGAACAGTGAGGTTATAC 3&#39; (SEQ ID NO: 6)SalI coup-  End of D1ling sitePrimer (5&#39;)                          LAG-3 D1-D4identical to LAG-3 D1                from the leaderPrimer (3&#39;)                          sequence to5&#39; GCGCGTCGACTTAACCCTGGGCTAGACAGCTCTCTGTG 3&#39; (SEQ ID NO:                                amino acid 412SalI coup-  End of D4ling site__________________________________________________________________________ 
    
     The resulting amplified fragments were digested with SalI and inserted into the SalI site of pUC 18 (Stratagene). 
     The amplified sequences were verified, and the inserts subcloned into the expression vector pCLH3 AXS V2 DHFR hα IVS as described by COLE et al. (Biotechnology 11, 1014-1024, 1993) (FIG. 4). 
     This vector is a eukaryotic expression vector which is multifunctional for the expression CDNA and its amplification in eukaryotic cells. It possesses the following features: (i) the murine promoter of the metallothionein-1 gene and a polyadenylation sequence SV 40 (comprising a donor-acceptor splicing site) to bring about transcription of the gene of interest, (ii) a human intervening sequence A containing the donor-acceptor splicing site of the gene for the subunit of α glycoprotein for obtaining high levels of transcription of cDNA, (iii) the pML sequence containing the origin of replication of pBR 322 and a gene for resistance to aampicillin for bacterial amplification, and (iv) a DHFR transcription unit of SV 40 to bring about transcription of the sequences used for selection and amplification of the transfectants. 
     Stable expression in CHO cells 
     The expression vectors coding for LAG-3 D1, LAG-3 D1D2 and LAG-3 D1-D4 were used to transfect CHO DUKX cells, and these cells were cultured on a selective medium. Cells capable of multiplying under these conditions were combined and cultured on a medium containing increasing amounts of MTX. Levels of expression were measured by Western blot analysis using the 17B4 monoclonal antibody. Clones producing high levels of recombinant soluble molecules derived from LAG-3 were propagated in bioreactors, and the material derived from LAG-3 was purified by ion exchange chromatography and immunoaffinity. 
     Western blot analyses revealed, in supernatants of cells transfected with expression vectors coding for LAG-3 D1, LAG-3 D1D2 and LAG-3 D1-D4, bands with apparent Mr values of 15 to 18 kD, 34-36 kD (doublets) and 55 kD (2 possible bands) . The respective Mr values of these immunoreactive materials corresponded to the expected Mr values of glycosylated LAG-3 D1 Ig (139 amino acids and a putative N-glycosylation site), glycosylated LAG-3 D1D2 Ig (239 amino acids containing 3 glycosylation sites) and glycosylated LAG-3 D1-D4 Ig (412 amino acids containing 4 glycosylation sites). 
     EXAMPLE 4 
     Specific binding of LAG-3 Ig to cells expressing MHC Class II 
     The reactivity of the monoclonal antibodies and of LAG-3 D1-D4 Ig was studied by indirect immunofluorescence. Target cells (4×10 5 ) were incubated for 30 minutes at 4° C. in the presence of LAG-3 D1-D4 Ig, CD8 Ig, a murine monoclonal antibody, (949) anti-human MHC Class II (DR, DP, DQ) conjugated to FITC (isothiocyanate fluoride) from a Coulter clone, or murine Ig-FITC: an irrelevant immunoglobulin G conjugated to FITC. The cells were washed and incubated at 4° C. for 30 minutes with either a goat anti-human Ig polyclonal F(ab&#39;) 2  conjugated to fluorescein or a goat anti-mouse Ig polyclonal antibody conjugated to fluorescein (Coulter clone). 
     To confirm the LAG-3/MHC Class II binding, LAG-3 D1-D4 Ig was incubated with MHC Class II-positive or -negative cells. Four B lymphocyte lines expressing MHC Class II(L31, Phil EBV, Raji, Sanchez and Personnaz) were treated with anti-Class II monoclonal antibody 949, or the supernatants of Cos cells transfected with DNA coding either for LAG-3 D1-D4 Ig or for CD8 Ig. The five cell lines expressing the different haplotypes of MHC Class II molecules were recognized by LAG-3 Ig in the same way as by the anti-Class II monoclonal antibodies (positive control), while the supernatant containing CD8 Ig (negative control) did not bind to these cell lines, as could be expected. Four MHC Class II-negative cell lines (CEM, RJ, HSB2, K562) were treated with the same reagents as above. None reacted, either with the anti-MHC Class II (negative control) or with LAG-3 D1-D4 Ig, showing that the binding of LAG-3 D1-D4 is specific to MHC Class II molecules. 
     Further experiments were carried out using (i) mouse fibroblasts transfected or otherwise with genes coding for human DR7 or human DP4, (ii) mouse cells expressing or otherwise MHC Class II molecules, (iii) activated human CD4 +   or CD8 +   cells, and (iv) T lymphocyte lines expressing the different haplotypes of MHC Class II molecules (FIG. 8). 
     Unlike CD8 Ig, LAG-3 D1-D4 Ig binds to all cells expressing MHC Class II as efficiently as the anti-MHC Class II monoclonal antibody 949. LAG-3 D1-D4 Ig binds to all DR and DP haplotypes tested, to human MHC Class II molecules expressed by transfected mouse cells, to murine MHC Class II molecules and also to MHC Class II molecules expressed by CD4 +   or CD8 +   T lymphocytes. 
     These results represent for the first time proof that soluble molecules derived from a ligand for MHC Class II are capable of binding to cells expressing MHC Class II. 
     Similar experiments showed that LAG-3 D1D2 bound to cells expressing MHC Class II in as specific a manner and with the same efficiency as LAG-3 D1-D4. 
     Binding activity of LAG-3Ig and cellular distribution of ligands for LAG-3Ig 
     The capacity of this immunoadhesin to bind to cell ligands is measured using a fluorescein-labelled goat serum directed against human immunoglobulins. 
     In these experiments, the target cells are first incubated with a human monoclonal antibody or an immunoadhesin for 30 min at 4° C. in RPMI 1640 containing 10% of FCS (foetal calf serum). The cells are then incubated with an FITC-labelled goat anti-mouse immunoglobulin serum (Coulter) for the murine monoclonal antibodies or with an FITC-labelled goat anti-human immunoglobulin serum (Tago) for the immunoadhesins. The fluorescence is measured after two washes, analyzing 3,000 cells with an Elite cytometer (Coultronics, Hialeah, Fla.). FIG. 9 shows the degrees of binding of LAG-3Ig, CD8Ig, antibody 949 or antibody OKT3 (anti-CD3, ATCC), represented by the number of cells counted as a function of the logarithm of the measured fluorescence intensity. 
     LAG-3Ig binds to mouse fibroblasts transfected for the gene for the HLA DR 4  molecule, and does not bind to untransfected cells. CD8Ig is incapable of binding to HLA DR 4   +   fibroblasts under the same conditions. 
     The cellular distribution of the ligands for LAG-3Ig was evaluated on a cell population sample by immunofluorescence. 
     LAG-3Ig is visualized on all positive Class II cells tested, including B cell lines transformed by Epstein-Barr virus (derived from genetically unrelated donors, including 10 homozygous lines of DR 1  to DR 10  typing), as well as on activated T and NK cells. 
     FIG. 9 shows, by way of example, the binding of LAG-3Ig to Daudi cells which are positive for Class II antigens. 
     The mean fluorescence intensity with LAG-3Ig is similar to that observed with antibody 949 which is specific for Class II antigens. The binding of LAG-3Ig to DR 4  (FIG. 9), DR 2 , DR 7  or DPw4 (not shown) expressed at the surface of mouse fibroblasts is, in contrast, weaker than that observed for antibody 949. 
     No binding is detected on cell lines which are negative for Class II antigens of T origin (peripheral blood T cells, CEM, HSB2, REX lines), of B origin (RJ 2.2.5 line) or of non-lymphoid origin (human lines, K562 of erythromyoloid origin and line originating from melanoma cells (not shown)). 
     Moreover, LAG-3Ig binds to xenogeneic Class II molecules of the MHC, such as the antigens expressed by mouse lymphoid A 20 and the monkey Classes II expressed by phytohaemagglutinin-stimulated blasts (data not shown). 
     The specificity of binding of LAG-3Ig was also verified using the monoclonal antibodies 17B4, whose capacity to block LAG-3/MHC Class II interactions in cell adhesion tests was demonstrated beforehand (FIG. 10). 
     In these experiments, the LAG-3Ig molecules are preincubated for 30 minutes at 4° C. either with medium alone, or with 17B4 (1 mg/ml), or with OKT3 (1 mg/ml), before being brought into contact with Daudi cells. 
     FIG. 10 shows that a preincubation of LAG-3Ig with 17B4 inhibits the binding to Class II +   cells, whereas no inhibition is detected with the OKT3 control. 
     EXAMPLE 5 
     Inhibition of LAG-3/MHC Class II interaction by soluble fragments of LAG-3 
     The inhibition of LAG-3/MHC Class II interaction by the soluble fragments of LAG-3 may be observed directly in relation to the binding of LAG-3Ig by Class II MHC molecules, by competitive experiments with the soluble fragments. 
     To verify whether the soluble LAG-3D 1  D 2  fragments produced by CHO cells could displace the binding of immunoadhesins derived from LAG-3, the following tests were carried out: 
     Daudi cells are incubated with soluble LAG3-D 1  D 2  fragments so as to permit the binding of these molecules to the MHC Class II antigens expressed at the surface of the Daudi cells. 
     In a second step, the cells are incubated in the presence of LAG-3D 1  D 4  Ig in dimeric form or LAG-3D 1  D 2  Ig in monomeric form. 
     The binding of these immunoadhesins derived from LAG-3 is measured using a goat anti-human Ig F(ab&#39;) 2  conjugated to fluorescein (GAH-FITC). 
     The control groups are represented by Daudi cells incubated with dimeric LAG-3D 1  D 4  Ig or monomeric LAG-3D 1  D 2  Ig without preincubation with the soluble LAG-3D1D2 fragments. 
     The results are recorded in Table 5, which shows that the soluble LAG-3D 1  D 2  fragments are capable of displacing the immunoadhesins derived from LAG-3 in monoor dimeric form. 
     
                       TABLE 5______________________________________                Mean Fluor-Reactants  Detection escence   Conclusion______________________________________--         GAH-FITC  0.3       GAH does not                          interfereDimeric    GAH-FITC  20.8      The binding ofLAG-3D1D4Ig                    CHO/LAG-3D1D2                          inhibits the bindingCHO/LAG-3D1D2,      GAH-FITC  8.5       of dimericthen dimeric                   LAG-3D1D4Ig (58%)LAG-3D1D4IgMonomeric  GAH-FITC  62.5      The binding ofLAG-3D1D2Ig                    CHO/LAG-3D1D2                          inhibits the bindingCHO/LAG-3D1D2,      GAH-FITC  10.9      of monomericthen monomeric                 LAG-3D1D2Ig (27%)LAG-3D1D2Ig______________________________________ 
    
     These data confirm that the soluble LAG-3D1D2 fragments bind to MHC Class II molecules. 
     Inhibition of LAG-3/MHC Class II and CD4/MHC Class II interaction 
     Rosette formation between Cos cells transfected with wild-type LAG-3 and B lymphocytes transformed with EBV expressing MHC Class II molecules was demonstrated by BAIXERAS et al. (2). This interaction is inhibited both by anti-LAG-3 and anti-MHC Class II monoclonal antibodies. 
     The method described in this publication was modified by replacing the visualization and counting of Cos cells binding to B lymphocytes by counting the radioactivity remaining after incubation of  51  Cr-labelled B lymphocytes with Cos cells expressing LAG-3 (binding assay). 
     The possible inhibitory effects of soluble molecules derived from LAG-3 on LAG-3/MHC Class II interaction, and also on CD4/MHC Class II interaction, were studied. 
     Cos cells transfected with a suitable expression vector (coding for wild-type LAG-3 or for CD4). Two days later, the Cos cells were treated with trypsin and plated out again on the basis of 0.05×106 cells/well on flat-bottomed 12-well tissue culture plates. 24 hours later,  51  Cr-labelled Daudi cells (5.5×10 6 ) were incubated on this monolayer of Cos cells (final vol.: 1 ml) for 1 hour. The target B cells were then aspirated off and the wells washed 5 to 7 times, gently adding 1 ml of medium dropwise. The edges of the wells were washed by suction using a Pasteur pipette. The remaining cells were lysed with 1 ml of PBS, 1% Triton for 15 minutes at 37° C. The lysates were centrifuged at 3000 rpm for 10 minutes, and 100 μl of the resulting supernatant were counted. 
     LAG-3 D1-D4 Ig was used to inhibit LAG-3/MHC Class II and CD4/MHC Class II interaction in the  51  Cr binding assay. Human CD8 Ig and IgG1 were tested in parallel and used as negative controls. 
     A significant inhibition of LAG-3/Class II interaction by LAG-3 D1-D4 Ig was detected (FIG. 5A). However, the LAG-3/MHC Class II interaction can be partially and non-specifically inhibited by human CD8 Ig and IgG1. Moreover, LAG-3 Ig proved to be a potential inhibitor of CD4/Class II interaction (FIG. 5B) under experimental conditions in which CD4/MHC Class II interaction was not modified by human CD8 Ig or IgG1. This suggests that LAG-3/Class II interaction is weaker than CD4/Class II interaction. These results represent the first proof of a possible competition of soluble molecules in an interaction of MHC Class II with its ligands. 
     EXAMPLE 6 
     Immunosuppressant activity of LAG-3 D1-D4 Ig 
     Functional tests were performed using the proliferation tests described above for the biological activity of the anti-LAG-3 monoclonal antibodies. 
     3 days and 5 days (D3 and D5) after antigenic stimulation, LAG-3 D1-D4 Ig showed a strong inhibition of the proliferation of clone 28, while human CD8 Ig and IgG had no effect (FIG. 6). Similar experiments were carried out with clone 154 (FIG. 7), and showed a partial inhibition in the presence of LAG-3 Ig. A control carried out with anti-LAG-3 monoclonal antibodies had the reverse effects, as observed previously. 
     A significant inhibition of the cell proliferation of cells incubated in the presence of LAG-3 D1-D4 Ig was also observed for clone 28. 
     These observations show that LAG-3 D1-D4 Ig is a potential immunosuppressant of the proliferation of T lymphocytes stimulated by an antigen, and indicate that LAG-3 might act as an &#34;extinguisher&#34; of the secondary immune response induced by activated CD4 +   T helper lymphocytes. 
     Role of LAG-3Ig in the negative regulation of the immune responses of T cells 
     To demonstrate that a soluble form of LAG-3, mimicking the functions of the membrane molecule, could inhibit the activation of CD 4   +   T clones stimulated by an antigen, the following tests were carried out on clone T154: the T cells are incubated beforehand with a saturating amount of LAG-3Ig (100 nM). The cells are then washed twice with cold RPMI and incubated with 10 μg/ml of goat antibodies directed against human immunoglobulins (Tago) at 4° C. for 30 minutes. 
     After two more washes, the cells are resuspended in RPMI containing 10% of foetal calf serum and incubated for 2 hours at 37° C. before adding the signal. To couple (&#34;cross-link&#34;) the monoclonal antibodies, a goat anti-mouse antibody at a concentration of 10 μg/ml (Tago) is used. 
     FIG. 11 depicts an experiment in which clone T154 has been preincubated with LAG-3Ig bound (&#34;cross-linked&#34;) to a second reactant (polyclonal serum specific for the constant region of human immunoglobulins). The degree of binding of LAG-3Ig to the cells is measured by immunofluorescence (FIG. 11A). FIG. 11B shows that a more than 50% inhibition of the proliferation of clone T154 is produced by LAG-3Ig. Under the same experimental conditions, no effect is observed with the control CD8Ig or with LAG-3Ig without &#34;cross-linking&#34; (not shown in the figure). 
     FIG. 11C also shows that no effect is observed when LAG-3Ig is used to bind (&#34;cross-link&#34;) the MHC Class II molecules expressed by antigen-presenting B cells. 
     The possible effects of bound (&#34;cross-linked&#34;) anti-Class II monoclonal antibodies in relation to the proliferation of T cells were compared to those of LAG-3Ig. A weak inhibition (less than 50%) is observed with antibody 949 and antibody D1.12 (anti-DR) bound to a goat anti-mouse polyclonal serum (FIG. 12). The inhibition of proliferation is hence epitope-dependent, the largest effect being obtained with the epitope of LAG-3 specific for the binding to Classes II. 
     The effects of LAG-3 Ig on the proliferation of T cells were also studied using different signals on another CD 4   +   T clone, clone TDEL specific for peptide 34-53 of the basic myelin protein. 
     An inhibition of proliferation is observed (n=2) when TDEL is stimulated with the antigen (not shown), with immobilized OKT3 (FIG. 13A), with lectins (PHA+PMA) (FIG. 13B) and with 5 IU/ml of IL 2  (FIG. 13C). No inhibition is observed with 100 IU/ml of IL 2  (FIG. 13D). 
     In conclusion, these results collectively show that LAG-3 and MHC Class II molecules, which are each T cell-activating antigens, may be likened to effector molecules involved in the phase of inactivation of T cell responses. Moreover, these results illustrate the importance of interactions between T cells in the control of the cellular immune response. 
     EXAMPLE 7 
     Stimulation of cell cytotoxicity by LAG-3Ig 
     The role of LAG-3Ig in relation to cell cytotoxicity is studied on two types of effector cells: 
     freshly drawn human peripheral blood lymphocytes (PBL), 
     S1B5 line cells (clone of human NK cells). 
     The cytotoxic activity of these cells is measured by counting the  51  Cr released into the medium by previously labelled target cells, in the presence or absence of LAG-3Ig in the medium. 
     FIG. 14 shows the degree of cytotoxicity of S1B5 for a line of human B cells transformed by Epstein-Barr virus and carrying major histocompatibility complex Class I and II antigens (LAZ 388 line), as a function of different reactants added to the cultures. 
     Measurements are carried out after 4 hours of coculture for effector/target (S1B5/LAZ 388) cell ratios of 3:1 (clear columns) or 1:1 (shaded columns). 
     The negative controls consist of medium alone (MED), the immunoadhesin CD8Ig and the monoclonal antibody 17.B4 (anti-LAG-3). 
     The positive controls consist of three different monoclonal antibodies: 
     antibody L243 directed against Class II DR antigens, 
     antibody 9.49 directed against Class II DR, DP, DQ antigens, 
     antibody W632 directed against human major histocompatibility complex Class I antigens. 
     Anti-HLA Class I (W632) or Class II (L243) antibodies increase the lysis of the target cells (and not the 17B4 control). The immunoadhesin LAG-3Ig increases the lysis. The CD8Ig control has no effect. 
     FIG. 15 shows the results of an experiment similar to the above, in which the cytotoxicity of PBL with respect to Daudi cells (HLA Class I - ) is measured, for effector/target ratios of 50:1 (clear columns) and 15:1 (shaded columns). The reactants added to the medium are the same as the ones used in the first experiment, except for antibody 9.49 and antibody 17.B4. Antibody 10H3 is an isotype IgG1 immunoglobulin specific for the CD45 surface antigen. It is used as negative control. 
     No change is observed with an antibody directed against major histocompatibility complex Class I antigens (W632). 
     The data from these two series of measurements show that, compared to negative controls, LAG-3Ig activates the cytotoxicity of NK cells. This effect is similar to the one observed with antibodies directed against MHC Class II molecules. 
     BIBLIOGRAPHIC REFERENCES 
     1. TRIEBEL T. et al., 1990, J. Exp. Med. 171, 1393-1405 
     2. BAIXERAS E. et al., 1992, J. Exp. Med. 176, 327-337 
     3. COSGROVE D. et al., 1991, Cell 66, 1051-1066 
     4. RAHEMTULLA A. et al., 1991, Nature 353, 180-184 
     5. TRAUNECKER A. et al., 1988, Nature 331, 84-86 
     6. BENEDICT A. A. et al., 1967, Methods in Immunology 1, 197-306 (1967) 
     7. YELTON D. E. et al., Ann. Rev. of Biochem. 50, 657-680 (1981) 
     8. HUARD B. et al., Immunogenetics 39: 213 
     9. MANIATIS T. et al. (1982), Molecular cloning: A laboratory manual--Cold Spring Harbor Laboratory, New-York. 
     10. SEED B., 1987, Nature 329, 840-842 
     11. COLE S. C. et al., Biotechnology 11, 1014-1024, 1993 
     12. COLE S. C. et al., Biotechnology 11, 1014-1024, 1993. 
     
                                           TABLE NO. 1__________________________________________________________________________                    reisdue                          atom type,Atom                     name and                          charge andname    x      y      z      no.   no.__________________________________________________________________________N   25.172370911      27.259836197             67.855064392                    AP-n                        40                          n3                            -0.5000                                 1NN2 25.667764664      26.471963882             67.420585632                    AP-n                        40                          hn                             0.1300                                 2CA  24.625223160      26.867494583             69.180244446                    AP-n                        40                          ca                             0.1200                                 3NN1 24.393711090      27.474891663             67.220008850                    AP-n                        42                          hn                             0.1300                                 4MA  23.936895370      27.680395126             69.464080811                    AP-n                        40                          h  0.0700                                 5C   25.662780726      26.773513794             70.350120544                    AP-n                        40                          c&#39;                             0.3800                                 6O   25.295415878      27.090747833             71.482070923                    AP-n                        40                          o&#39;                            -0.4100                                 7CB  23.766021729      25.587018967             68.990669250                    AP-n                        40                          c2                            -0.2600                                 8MB1 23.060285568      25.72443695             68.152351379                    AP-n                        40                          h  0.0700                                 9MB2 24.413969040      24.744903564             68.686981201                    AP-n                        40                          h  0.0700                                 10CG  22.921775818      25.153419495             70.196960449                    AP-n                        40                          c-                             0.3400                                 11DD1 22.069602966      25.929233551             70.676017761                    AP-n                        40                          o-                            -0.5700                                 12DD2 23.115716934      24.009321213             70.667663574                    AP-n                        40                          o-                            -0.5700                                 13M   26.906179428      26.304588318             70.124969482                    SER 41                          n -0.5000                                 14CA  27.860145569      25.912786484             71.207519531                    SER 41                          ca                             0.1200                                 15HN  27.120641708      26.221813202             69.126319885                    SER 41                          hn                             0.2800                                 16MA  27.374551773      25.088045120             71.766326904                    SER 41                          h  0.1000                                 17C   28.252065659      27.005065918             72.271789551                    SER 41                          c&#39;                             0.3800                                 18O   27.987834930      28.200620651             72.115295410                    SER 41                          o&#39;                            -0.3800                                 19CB  29.083601227      25.298025131             70.494880676                    SER 41                          c2                            -0.1700                                 20MB1 28.786190033      24.599395752             69.690521240                    SER 41                          h  0.1000                                 21MB2 29.691858292      26.092912674             70.004081726                    SER 41                          h  0.1000                                 22OG  29.905221939      24.578149796             71.424118042                    SER 41                          oh                            -0.3800                                 23HG  30.662555695      24.231645584             70.939277649                    SER 41                          ho                             0.3500                                 24N   28.857526779      26.558052063             73.387054443                    GLY 42                          n -0.5000                                 25CA  29.199718475      27.440891266             74.535232544                    GLY 42                          cg                             0.0200                                 26MN  29.251720428      25.616510391             73.267890930                    GLY 42                          hn                             0.2800                                 27MA1 28.520187378      28.312047958             74.591857910                    GLY 42                          h  0.1000                                 28MA2 28.983697891      26.890144348             75.468444824                    GLY 42                          h  0.1000                                 29C   30.691051483      27.875793457             74.601707458                    GLY 42                          c&#39;                             0.3800                                 30O   31.504199982      27.026502609             74.980445862                    GLY 42                          o&#39;                            -0.3800                                 31N   31.113182068      29.132183075             74.266487122                    PRO 43                          n -0.4200                                 32CA  32.858349609      29.476200104             74.126792908                    PRO 43                          ca                             0.0600                                 33HA  33.096603394      28.605407715             73.708091736                    PRO 43                          h  0.1000                                 34CD  30.24075089      30.174203873             73.722633362                    PRO 43                          c2                             0.0600                                 35MD1 29.467987061      30.516298294             74.466743469                    PRO 43                          h  0.1000                                 36MD2 29.664882660      29.799777985             72.838768005                    PRO 43                          h  0.1000                                 37C   33.318023682      29.988374710             75.414916992                    PRO 43                          c&#39;                             0.3800                                 38O   32.682361603      30.557033539             76.312332153                    PRO 43                          o&#39;                            -0.3800                                 39CB  32.483139038      30.574260712             73.043136597                    PRO 43                          c2                            -0.2000                                 40MB1 33.350620270      31.263189316             73.049049377                    PRO 43                          h  0.1000                                 41MB2 32.463790894      30.110534668             72.036743164                    PRO 43                          h  0.1000                                 42CG  31.160949707      31.299722672             73.307487488                    PRO 43                          c2                            -0.2000                                 43MG1 31.279897690      32.024162292             74.137779236                    PRO 43                          h  0.1000                                 44MG2 30.794561386      31.862808228             72.428352356                    PRO 43                          h  0.1000                                 45K   34.683673859      29.902477264             75.503486633                    PRO 44                          n -0.4200                                 46CA  35.485736847      30.679145813             76.490524292                    PRO 44                          ca                             0.0600                                 47KA  35.018527985      30.645456314             77.491592407                    PRO 44                          h  0.1000                                 48CD  35.509281158      29.014368057             74.655700684                    PRO 44                          c2                             0.0600                                 49MD1 35.411357880      29.247959137             73.577461243                    PRO 44                          h  0.1000                                 50MD2 35.214973450      27.955932617             74.801994324                    PRO 44                          h  0.1000                                 51C   35.700843811      32.172924042             76.063224792                    PRO 44                          c&#39;                             0.3800                                 52O   36.448230743      32.477428436             75.126922607                    PRO 44                          o&#39;                            -0.3800                                 53CB  36.779544830      29.842718124             76.547348022                    PRO 44                          c2                            -0.2000                                 54HB1 37.662769315      30.430650711             75.863670149                    PRO 44                          h  0.1000                                 55HB2 36.667564392      29.027103424             77.288825989                    PRO 44                          h  0.1000                                 56CG  36.940769196      29.250995636             75.143180867                    PRO 44                          c2                            -0.2000                                 57MG1 37.446662903      29.982709885             74.483322144                    PRO 44                          h  0.1000                                 58MG2 37.553295135      26.329860667             75.134750366                    PRO 44                          h  0.1000                                 59N   35.026676178      33.104183197             76.753837585                    ALA 45                          h -0.5000                                 60CA  35.034278870      34.544979095             76.400695801                    ALA 45                          ca                             0.1200                                 61MN  34.452354431      32.747509003             77.536170959                    ALA 45                          hn                             0.2800                                 62MA  35.105010986      34.659946442             75.298950195                    ALA 45                          h  0.1000                                 63C   36.209384918      35.322113037             77.076705833                    ALA 45                          c&#39;                             0.3800                                 64O   36.163528442      35.637268066             78.268325806                    ALA 45                          o&#39;                            -0.3800                                 65CB  33.646369934      35.083190918             76.800727844                    ALA 45                          c3                            -0.3000                                 66MB1 33.534698486      36.150661469             76.535202026                    ALA 45                          h  0.1000                                 67MB2 32.828392029      34.539138794             76.290328979                    ALA 45                          h  0.1000                                 68MB3 33.465579987      35.001335144             77.890525818                    ALA 45                          h  0.1000                                 69N   37.266757965      35.613758087             76.297294617                    ALA 46                          n -0.5000                                 70MN  37.216701508      35.231555939             75.346885681                    ALA 46                          hn                             0.2800                                 71CA  38.489383698      36.310386658             76.786270142                    ALA 46                          ca                             0.1200                                 72MA  38.262126923      36.871456146             77.716934204                    ALA 46                          h  0.1000                                 73C   39.058414459      37.311935425             75.727844238                    ALA 46                          c&#39;                             0.3800                                 74O   38.922710419      37.100418091             74.516731262                    ALA 46                          o&#39;                            -0.3800                                 75CB  39.526046753      35.215301514             77.108406067                    ALA 46                          c3                            -0.3000                                 76MB1 40.446556091      35.633434296             77.555480957                    ALA 46                          h  0.1000                                 77MB2 39.131206512      34.469978333             77.822120667                    ALA 46                          h  0.1000                                 78MB3 39.821903229      34.663463593             76.197715759                    ALA 46                          h  0.1000                                 79N   39.737388611      38.384365082             76.180320740                    ALA 47                          n -0.5000                                 80CA  40.295833588      39.429889679             75.275863647                    ALA 47                          ca                             0.1200                                 81MN  39.717037201      38.512592316             77.196357727                    ALA 47                          hn                             0.2800                                 82MA  39.901103973      39.319335938             74.245994568                    ALA 47                          h  0.1000                                 83C   41.869789124      39.413467407             75.170806885                    ALA 47                          c&#39;                             0.3800                                 84O   42.518333435      40.166862488             75.906578044                    ALA 47                          o&#39;                            -0.3800                                 85CB  39.722030640      40.769996643             75.786285400                    ALA 47                          c3                            -0.3000                                 86MB1 40.045078278      41.611721039             75.145561218                    ALA 47                          h  0.1000                                 87MB2 38.615306854      40.778987885             75.787467957                    ALA 47                          h  0.1000                                 88MB3 40.059757233      41.007873535             76.813346863                    ALA 47                          h  0.1000                                 89N   42.537422180      38.621597290             74.274406433                    PRO 48                       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HIS 57                          h  0.1000                                214CB  43.250400543      33.671833038             75.751731873                    HIS 57                          c5                             0.1000                                215MD1 44.116428375      33.723646257             74.666069031                    HIS 57                          np                            -0.4200                                216CE1 43.325973511      33.139041901             73.713264465                    HIS 57                          c5                             0.2700                                217ME2 42.066158295      32.716590881             74.036811829                    HIS 57                          np                            -0.5000                                218CD2 42.045005798      33.048488617             75.379264832                    HIS 57                          c5                             0.0100                                219ME1 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62                          hn                             0.2800                                273MA  37.921787262      35.006183624             79.841125488                    SER 62                          h  0.1000                                274C   37.145660400      33.532897949             81.261909485                    SER 62                          c&#39;                             0.3800                                275O   37.466091156      32.626064301             82.041137695                    SER 62                          o&#39;                            -0.3800                                276CB  39.620265961      34.048240662             80.725196838                    SER 62                          c2                            -0.1700                                277MB1 39.660293579      33.306644440             79.904281616                    SER 62                          h  0.1000                                278MB2 40.349685669      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               284MA  35.064254761      32.265518188             82.170570374                    SER 63                          h  0.1000                                285C   34.106758118      31.936998367             80.231674194                    SER 63                          c&#39;                             0.3800                                286O   33.716896057      32.367130280             79.142120361                    SER 63                          o&#39;                            -0.3800                                287CB  33.716815948      33.889484406             81.843544006                    SER 63                          c2                            -0.1700                                288MB1 34.199871063      34.571384430             82.572105408                    SER 63                          h  0.1000                                289MB2 33.32880719      34.543502808             81.036796570                    SER 63                          h  0.1000                                290OG  32.634590149      33.222091675             82.496467590                    SER 63                          oh                            -0.3800                                291MG  32.159793854      32.710407257             81.832328796                    SER 63                          ho                             0.3500                                292N   33.914913177      30.658897400             80.588378906                    TRP 64                          n -0.5000                                293CA  33.112319946      29.694124222             79.783546448                    TRP 64                          ca                             0.1200                                294MN  34.221500397      30.422899246             81.538955688                    TRP 64                          hn                             0.2800                                295MA  33.404731750      29.812168121             78.721931458                    TRP 64                          h  0.1000                                296C   31.573041916      29.961977005             79.883049011                    TRP 64                          c&#39;                             0.3800                                297O   30.996980667      29.940891266             80.975959778                    TRP 64                          o&#39;                            -0.3800                                298CB  33.525466919      26.235471725             80.142707825                    TRP 64                          c2                            -0.2000                                299MB1 32.950366974      27.534402847             79.508041382                    TRP 64                          h  0.1000                                300MB2 34.571674347      28.078489304             79.816658020                    TRP 64                          h  0.1000                                301CG  33.405326843      27.783784866             81.611763000                    TRP 64                          c5                             0.0000                                302CD1 32.267101288      27.214570999             82.221214294                    TRP 64                          c5                             0.1000                                303NE1 32.481933594      26.953943253             83.590408325                    TRP 64                          np                            -0.5000                                304CE2 33.781982422      27.378627777             83.812339783                    TRP 64                          c5                             0.1100                                305CD2 34.355152130      27.881061554             82.617965698                    TRP 64                          c5                             0.0000                                306MD1 31.322025528      27.036033630             81.708480835                    TRP 64                          h  0.1000                                307ME1 31.820940018      26.578481674             84.279945374                    TRP 64                          hn                             0.2800                                308CE3 35.681034088      28.394184113             82.615905762                    TRP 64                          cp                            -0.1000                                309ME3 36.128986359      28.784986496             81.714393616                    TRP 64                          h  0.1000                                310CE3 36.396430969      28.387191772             83.815704346                    TRP 64                          cp                            -0.1000                                311ME3 37.405311584      28.776082993             83.832160950                    TRP 64                          h  0.1000                                312CM2 35.830604553      27.888952255             84.994010925                    TRP 64                          cp                            -0.1000                                313MM2 36.410472870      27.896844844             85.906951904                    TRP 64                          h  0.1000                                314CZ2 34.527233124      27.382776260             85.014289856                    TRP 64                          cp                            -0.1000                                315MZ2 34.097515106      27.006088257             85.929389954                    TRP 64                          h  0.1000                                316N   30.921349498      30.232547760             78.740600586                    GLY 65                          n -0.5000                                317CA  29.460748672      30.504768372             78.692192078                    GLY 65                          cg                             0.0200                                318MN  31.520374298      30.302478790             77.901519775                    GLY 65                          hn                             0.2800                                319MA1 29.073087692      30.896234512             79.650825500                    GLY 65                          h  0.1000                                320MA2 29.288171768      31.333106995             77.981094360                    GLY 65                          h  0.1000                                321C   28.633579254      29.293350220             78.197364807                    GLY 65                          c&#39;                             0.3800                                322O   28.866907883      29.137302399             76.975486755                    GLY 65                          o&#39;                            -0.3800                                323N   27.969013672      28.429246902             79.038352966                    PRO 66                          n -0.4200                                324CA  27.282257080      27.212890625             78.846752930                    PRO 66                          ca                             0.0600                                325MA  27.989650726      26.634012222             77.917152405                    PRO 66                          h  0.1000                                326CD  28.016592026      28.832529831             80.511337280                    PRO 66                          c2                             0.0600                                327MD1 27.731332779      29.836006927             80.880874634                    PRO 66                          h  0.1000                                328MD2 29.027824402      28.301166534             80.897590637                    PRO 66                          h  0.1000                                329C   25.977466583      27.479314804             77.725341797                    PRO 66                          c&#39;                             0.3800                                330O   25.217950821      28.417282104             77.982574463                    PRO 66                          o&#39;                            -0.3800                                331CB  27.045602798      26.422634125             79.851196289                    PRO 66                          c2                            -0.2000                                332MB1 26.132562637      25.797079086             79.827728271                    PRO 66                          h  0.1000                                333MB2 27.890687943      25.729280472             80.029365540                    PRO 66                          h -0.1000                                334CG  27.003501892      27.477243423             80.958015442                    PRO 66                          c2                            -0.2000                                335MG1 25.990892410      27.921483994             81.014678955                    PRO 66                          h  0.1000                                336MH2 27.232566833      27.061700821             81.956459045                    PRO 66                          h  0.1000                                337N   25.734319687      26.626403809             76.719802856                    ARG+                        67                          n -0.5000                                338CA  24.603988647      26.793025970             75.767257690                    ARG+                        67                          ca                             0.1200                                339MN  26.386735916      25.841371536             76.649803162                    ARG+                        67                          hn                             0.2800                                340MA  24.496238708      27.874828339             75.561668396                    ARG+                        67                          h  0.1000                                341C   23.227464676      26.224872589             76.267372131                    ARG+                        67                          c&#39;                             0.2800                                342O   23.178310394      25.034952164             76.603759766                    ARG+                        67                          o&#39;                            -0.3800                                343CB  24.990058899      26.165229797             74.398826599                    ARG+                        67                          c2                            -0.2000                                344MB1 24.135663986      26.318639755             73.709175110                    ARG+                        67                          h  0.1100                                345MB2 25.787433624      26.779323578             73.940032959                    ARG+                        67                          h  0.1100                                346CG  25.439929962      24.676465988             74.361564636                    ARG+                        67                          c2                            -0.2000                                347MG1 26.546255112      24.646316528             74.415458679                    ARG+                        67                          h  0.1300                                348MG2 25.092346191      24.131168365             75.261718750                    ARG+                        67                          h  0.1300                                349CD  24.934387207      23.941221237             73.112297058                    ARG+                        67                          c2                            -0.0900                                350MD  23.838283539      23.774566650             73.188652039                    ARG+                        67                          h  0.1300                                351MD2 25.070211411      24.585262299             72.220893860                    ARG+                        67                          h  0.1300                                352ME  25.665744781      22.657058716             72.968780518                    ARG+                        67                          n1                            -0.5000                                353ME  26.251846313      22.313375473             73.731925964                    ARG+                        67                          hn                             0.3600                                354CZ  25.689014435      21.902687073             71.871635437                    ARG+                        67                          cr                             0.4500                                355MM1 26.493299484      20.879484177             71.859733582                    ARG+                        67                          n2                            -0.5000                                356M11 27.072875705      20.740955353             72.690315247                    ARG+                        67                          hn                             0.3600                                357MM12    26.520929337      20.119580078             71.006805420                    ARG+                        67                          hn                             0.3600                                358MM2 24.956668854      22.117029190             70.805320740                    ARG+                        67                          n2                            -0.5000                                359MM21    25.030595779      21.456792831             70.033142090                    ARG+                        67                          hn                             0.3600                                360MM22    24.266489029      22.916227341             70.850784302                    ARG+                        67                          hn                             0.3600                                361N   22.080270767      26.971176147             76.244255066                    ARG+                        68                          n -0.4200                                362CA  20.743734360      24.358839035             76.485237122                    PRO 68                          ca                             0.0600                                363MA  20.817556381      25.605155945             77.294357300                    PRO 68                          h  0.1000                                364CD  22.076143265      28.448001862             76.342918396                    PRO 68                          c2                             0.0600                                365MD1 22.539228439      28.949869156             75.469612122                    PRO 68                          h  0.1000                                366MD2 22.632146835      28.776126862             77.244499207                    PRO 68                          h  0.1000                                367C   20.182382584      25.586324692             75.240180969                    PRO 68                          c&#39;                             0.3800                                368O   20.420539856      24.381649017             75.139877319                    PRO 68                          o&#39;                            -0.3800                                369CB  19.948141098      27.549791336             77.062515259                    PRO 68                          c2                            -0.2000                                370MB1 18.858430862      27.490163803             76.877128601                    PRO 68                          h  0.1000                                371MB2 20.066789627      27.567596436             78.163002014                    PRO 68                          h  0.1000                                372CG  20.592071533      28.804227829             76.667758179                    PRO 68                          c2                            -0.2000                                373MG1 20.158363342      29.031393051             75.478172302                    PRO 68                          h  0.1000                                374MG2 20.428398132      29.704229355             77.092338562                    PRO 68                          h  0.1000                                375N   19.458055496      26.229068756             74.300010681                    ARG+                        69                          n -0.5000                                376CA  18.893756866      25.542047501             73.096710205                    ARG+                        69                          ca                             0.1200                                377MN  19.221296310      27.198928833             74.529014587                    ARG+                        69                          hn                             0.2800                                378MA  19.667610168      24.872461319             72.660797119                    ARG+                        69                          h  0.1000                                379C   18.514461517      26.608341217             72.012504578                    ARG+                        69                          c&#39;                             0.3800                                380O   17.383218765      27.099323273             72.036094666                    ARG+                        69                          o&#39;                            -0.3800                                381CB  17.681777954      24.654710770             73.539657593                    ARG+                        69                          c2                            -0.2000                                382MB1 18.011884689      23.935596466             74.315147400                    ARG+                        69                          h  0.1100                                383MB2 16.954420090      25.310214996             74.061965942                    ARG+                        69                          h  0.100                                384CG  16.946891785      23.862810135             72.427490234                    ARG+                        69                          c2                            -0.2000                                385MG1 16.649517059      24.851628113             71.612152100                    ARG+                        69                          h  0.1300                                386MG2 17.638776779      23.127803802             71.968805054                    ARG+                        69                          h  0.1300                                387CD  15.688191414      23.166488647             72.975959778                    ARG+                        69                          c2                            -0.0900                                388MD1 15.980383873      22.365550995             73.686569214                    ARG+                        69                          h  0.1300                                389MD2 15.090404510      23.898859024             73.554351807                    ARG+                        69                          h  0.1300                                390ME  14.889810562      22.612804413             71.848815918                    ARG+                        69                          n1                            -0.5000                                391ME  15.272388458      22.616128922             70.898582458                    ARG+                        69                          hn                             0.3600                                392CZ  13.644455910      22.143890381             71.942466736                    ARG+                        69                          cr                             0.4500                                393MM1 13.048615456      21.755460739             70.850708008                    ARG+                        69                          n2                            -0.5000                                394MM11    13.576630592      21.832328796             69.979103088                    ARG+                        69                          hn                             0.3600                                395MM12    12.090744019      21.411947250             70.936080933                    ARG+                        69                          hn                             0.3600                                396MM2 12.989639282      22.056533813             73.074882507                    ARG+                        69                          n2                            -0.5000                                397CZ  13.644455910      22.143890381             71.942466736                    ARG+                        69                          cr                             0.4500                                393MM1 13.048615456      21.755460739             70.850708008                    ARG+                        69                          n2                            -0.5000                                394MM11    13.576630592      21.822328796             69.979103088                    ARG+                        69                          hn                             0.3600                                395MM12    12.090744019      21.411947250             70.936080933                    ARG+                        69                          hn                             0.3600                                396MM2 12.989639282      22.056533813             73.074882507                    ARG+                        69                          n2                            -0.5000                                397MM21    12.033639908      21.696094513             73.066261292                    ARG+                        69                          hn                             0.3600                                398MM22    13.529273987      22.372974396             73.883934021                    ARG+                        69                          hn                             0.3600                                399N   19.436628342      26.928932190             71.074501038                    ARG+                        70                          n -0.5000                                400CA  19.223009109      27.811206818             69.878326416                    ARG+                        70                          ca                             0.1200                                401MN  20.357131958      26.451332092             71.165985107                    ARG+                        70                          hn                             0.2800                                402MA  19.087514877      27.065124512             69.071128845                    ARG+                        70                          h  0.1000                                403C   20.512538910      28.575149536             69.398536682                    ARG+                        70                          c&#39;                             0.3800                                404O   20.872812271      28.468791962             68.228363037                    ARG+                        70                          o&#39;                            -0.3800                                405CB  17.935552597      28.697717667             69.732887268                    ARG+                        70                          c2                            -0.2000                                406MB1 17.889257431      29.085472107             68.694030762                    ARG+                        70                          h  0.1100                                407MB2 17.053388596      28.033058167             69.807891846                    ARG+                        70                          h  0.1100                                408CG  17.775762558      29.883768082             70.717842102                    ARG+                        70                          c2                            -0.2000                                409MG1 18.004568100      29.946030045             71.747383118                    ARG+                        70                          h  0.1300                                410MG2 18.540782928      30.651863098             70.495643616                    ARG+                        70                          h  0.1300                                411CD  16.368116379      30.502414703             70.693214417                    ARG+                        70                          c2                            -0.0900                                412MD1 16.095691681      30.812221527             69.664886475                    ARG+                        70                          h  0.1300                                413MD2 15.630161285      29.711160660             70.937812805                    ARG+                        70                          h  0.1300                                414ME  16.255954742      31.590759277             71.711013794                    ARG+                        70                          n1                            -0.5000                                415ME  16.253637314      31.350564957             72.706428528                    ARG+                        70                          hn                             0.3600                                416CE  16.144437790      32.900745392             71.464561462                    ARG+                        70                          cr                             0.4500                                417MM1 16.055492401      33.712890625             72.481109619                    ARG+                        70                          n2                            -0.5000                                418MM11    16.071464539      33.294216156             73.413330078                    ARG+                        70                          hn                             0.3600                                419MM12    15.975571632      34.708621979             72.277374268                    ARG+                        70                          hn                             0.3600                                420MM2 16.120351791      33.420074463             70.259872437                    ARG+                        70                          n2                            -0.5000                                421MM21    16.025018692      34.432674408             70.167800903                    ARG+                        70                          hn                             0.3600                                422MM22    16.187112808      32.736862183             69.505996704                    ARG+                        70                          hn                             0.3600                                423N   21.115812302      29.562902451             70.071769714                    TYRC                        71                          n -0.5000                                424MN  21.515977859      30.157514572             69.338050842                    TYRC                        71                          hn                             0.2800                                425CA  22.034273148      29.314456940             71.218978882                    TYRC                        71                          ca                             0.1200                                426MA  22.671009064      28.444923401             70.976676941                    TYRC                        71                          h  0.1000                                427C   21.352920532      28.953493118             72.563385010                    TYRC                        71                          c&#39;                             0.4100                                428OCT 20.392858505      29.553161621             73.048652649                    TYRC                        71                          o&#39;                            -0.3800                                429O   21.928325653      27.853042603             73.145797729                    TYRC                        71                          oh                            -0.3800                                430MO  21.429273605      27.558662415             73.909782410                    TYRC                        71                          ho                             0.3500                                431CB  22.969152451      30.555358887             71.361984253                    TYRC                        71                          c2                            -0.2000                                432MB1 22.368267059      31.487627029             71.355415344                    TYRC                        71                          h  0.1000                                433MB2 23.401992798      30.559690475             72.381835938                    TYRC                        71                          h  0.1000                                434CG  24.144546509      30.666173935             70.352111816                    TYRC                        71                          cp                             0.0000                                435CD1 23.927837372      31.126276016             69.044418335                    TYRC                        71                          cp                            -0.1000                                436MD1 22.944635391      31.424867630             68.717597961                    TYRC                        71                          h  0.1000                                437CE1 24.987041473      31.212265015             68.143028259                    TYRC                        71                          cp                            -0.1000                                438ME1 24.819047928      31.555503845             67.131973267                    TYRC                        71                          h  0.1000                                439CZ  26.273118973      30.861675262             68.542274475                    TYRC                        71                          cp                             0.0300                                440OM  27.314481735      30.957365036             67.652267456                    TYRC                        71                          oh                            -0.3800                                441MN  26.980180740      31.236070633             66.796859741                    TYRC                        71                          ho                             0.3500                                442CE2 26.504697800      30.422637939             69.841377258                    TYRC                        71                          cp                            -0.1000                                443ME2 27.503232956      30.148866653             70.140815735                    TYRC                        71                          h  0.1000                                444CD2 25.447391510      30.325349808             70.743820190                    TYRC                        71                          cp                            -0.1000                                445MD2 25.652494431      29.968608856             71.745338440                    TYRC                        71                          h  0.1000                                446__________________________________________________________________________ *The numbering of the amino acids is shifted by minus 6 relative to the sequence SEQ ID No. 1. 
    
     
                                           TABLE NO. 2__________________________________________________________________________                    residue                          atom type,Atom                     name and                          charge andname    x      y      z      no.   no.__________________________________________________________________________N   24.753730401      26.435615520             68.246300842                    CYSn                        40                          nj                            -0.5000                                1CA   24.503000259      26.356292725             69.707687378                    CYSn                        40                          ca                            0.1200                                2NN1 23.861560822      26.429992676             67.734397888                    CYSn                        40                          hn                            0.1400                                3NN2 25.250690460      25.603437424             67.909477234                    CYSn                        40                          hn                            0.1400                                4NA   23.890571594      27.247760773             69.940788269                    CYSn                        40                          h 0.100                                5C     25.747190475      26.505004883             70.632949829                    CYSn                        40                          c&#39;                            0.3800                                6O     25.611124039      27.204542160             71.634971619                    CYSn                        40                          o&#39;                            -0.3800                                7CB   23.602088928      25.125436783             69.979545593                    CYSn                        40                          c2                            -0.3000                                8HB1 22.555475225      25.378625870             69.716011047                    CYSn                        40                          h 0.1000                                9HB2 23.865217209      24.277284622             69.317871094                    CYSn                        40                          h 0.1000                                10SG   23.613842010      24.466741562             71.679084778                    CYSn                        40                          s1                            0.1000                                11N     26.910152435      25.881416321             70.350471497                    SER 41                          n -0.5000                                12CA   28.052598953      25.721915930             71.310394287                    SET 41                          ca                            0.1200                                13HN   26.921674728      25.473436356             69.412322998                    SER 41                          hn                            0.2800                                14RA   27.761577606      24.886217117             71.978584290                    SER 41                          h 0.1000                                15C     28.477056503      26.929338455             72.226699829                    SER 41                          c&#39;                            0.3800                                16O     28.412267685      28.097608566             71.829902649                    SER 41                          o&#39;                            -0.3800                                17CB   28.257335662      25.212779999             70.480110168                    SER 41                          c2                            -0.1700                                18KB1 28.957101822      24.401222229             69.786743164                    SER 41                          h 0.1000                                19KB2 29.657680511      26.030597687             69.845893860                    SER 41                          h 0.1000                                20CG   30.284814825      24.713315964             71.339111328                    SER 41                          ch                            -0.3800                                21NG   31.027490616      24.448732376             70.785003662                    SER 41                          hc                            0.3500                                22N     28.904022217      24.617259979             73.466476440                    GLY 42                          n -0.5000                                23CA   29.226711273      27.639400482             74.497131348                    GLY 42                          cg                            0.0200                                24HX   29.140472412      25.626163483             73.587532043                    GLY 42                          hn                            0.2800                                25KA1 28.567264557      28.519987106             74.394309988                    GLY 42                          h 0.1000                                26KA2 28.949186325      27.229663849             75.483924866                    GLY 42                          h 0.1000                                27C     30.728670120      28.040773392             74.587509155                    GLY 42                          c&#39;                            0.3800                                28O     31.522300720      27.171119690             74.961143494                    GLY 42                          o&#39;                            -0.3800                                29N     31.175983429      29.296203613             74.282577515                    PRO 43                          n -0.4200                                30CA   32.627410889      29.612504959             74.140579224                    PRO 43                          ca                            0.0600                                31KA   33.153442383      28.711788635             73.723632813                    PRO 43                          h 0.1000                                32CO   30.295356750      30.374078751             73.784042358                    PRO 43                          c2                            0.0600                                33KD1 29.605636597      30.735929489             74.571166992                    PRO 43                          h 0.1000                                34KD2 29.683467865      30.826647568             72.928161621                    PRO 43                          h 0.1000                                35C     33.389945984      30.112844467             75.429031372                    PRO 43                          c&#39;                            0.3800                                36O     32.754360199      30.681560516             76.327354431                    PRO 43                          o&#39;                            -0.3800                                37CB   32.565906525      30.710325241             73.057388306                    PRO 43                          c2                            -0.2000                                38KB1 33.449081421      31.378034592             73.055488586                    PRO 43                          h 0.1000                                39KB2 32.524932861      30.247560501             72.051818848                    PRO 43                          h 0.1000                                40CG   31.263490677      31.466632843             73.332626343                    PRO 43                          c2                            -0.2000                                41KG1 31.413110733      32.204200745             74.146759033                    PRO 43                          h 0.1000                                42KG2 30.894020081      32.022109985             72.450286865                    PRO 43                          h 0.1000                                43N     34.754848480      30.015562057             75.523460388                    PRO 44                          n -0.4200                                44CA   35.553565979      30.763086319             76.536285400                    PRO 44                          ca                            0.0600                                45KA   35.083564758      30.695350647             77.536567688                    PRO 44                          h 0.1000                                46CD   35.574893951      29.225835648             74.665214519                    PRO 44                          c2                            0.0600                                47KD1 35.471595764      29.373666763             73.588935852                    PRO 44                          h 0.1000                                48KD2 35.281650543      28.076414108             74.807395935                    PRO 44                          h 0.1000                                49C     35.767509460      32.265411377             76.141113281                    PRO 44                          c&#39;                            0.3300                                50O     36.544441223      32.599441528             75.238464355                    PRO 44                          c&#39;                            0.3800                                51CB   36.849227905      29.927103043             76.567779541                    PRO 44                          c2                            -0.2000                                52HB1 37.732776642      30.502979279             76.899009705                    PRO 44                          h 0.1000                                53HB2 36.733722687      29.095364598             72.253833069                    PRO 44                          h 0.1000                                54CG   37.005489349      29.369808197             75.152618408                    PRO 44                          c2                            -0.2000                                55MG1 37.502185822      30.119005203             74.504158020                    PRO 44                          h 0.1000                                56MG2 37.625408173      28.451385498             75.115615845                    PRO 44                          h 0.1000                                57N     35.047088623      33.173435211             76.816978455                    ALA 45                          n -0.5000                                58CA   35.011333466      34.609920502             76.449218750                    ALA 45                          ca                            0.1200                                59MN   34.471405029      32.798248291             77.590728760                    ALA 45                          hn                            0.2800                                60HA   35.065380096      34.699813843             75.343650818                    ALA 45                          h 0.1000                                61C     36.187728882      35.414947510             77.090148926                    ALA 45                          c&#39;                            0.3800                                62O     36.133388519      35.819305420             78.255142212                    ALA 45                          o&#39;                            -0.3800                                63CB   33.615478516      35.135440826             76.831176758                    ALA 45                          c3                            -0.3000                                64HB1 33.490375519      36.187480927             76.517280579                    ALA 45                          h 0.1000                                65HB2 32.811222076      34.556259155             76.338432312                    ALA 45                          h 0.1000                                66HB3 33.433517456      35.098365784             77.922439575                    ALA 45                          h 0.1000                                67N     37.264499664      35.613868713             76.306388855                    ALA 46                          n -0.5000                                68HN   37.248832703      35.072978973             75.433502197                    ALA 46                          hn                            0.2800                                69CA   38.503662109      36.298694611             76.764076233                    ALA 46                          ca                            0.1200                                70MA   38.303600311      36.883266449             77.688095093                    ALA 46                          h 0.1000                                71C     39.082061768      37.273509979             75.687866211                    ALA 46                          c&#39;                            0.3800                                72O     38.951850891      37.052509308             74.481193542                    ALA 46                          c&#39;                            -0.3800                                73CB   39.509185791      35.179004669             77.103065491                    ALA 46                          c3                            -0.3000                                74HB1 40.441535950      35.582756042             77.525072327                    ALA 46                          h 0.1000                                75HB2 39.106670380      34.460605621             77.839447021                    ALA 46                          h 0.1000                                76HB3 39.780502319      34.997728729             76.205062866                    ALA 46                          h 0.1000                                77N     39.768814087      38.344066620             76.133750916                    ALA 47                          n 0.5000                                78CA   40.322643280      39.391151428             75.225708008                    ALA 47                          ca                            0.1200                                79MN   39.783836365      38.455593109             77.149337769                    ALA 47                          hn                            0.2800                                80HA   39.932807922      39.265201569             74.196365356                    ALA 47                          h 0.1000                                81C     41.900882721      39.401574817             75.126907349                    ALA 47                          c&#39;                            0.3800                                82O     42.538444519      40.171451569             75.854652405                    ALA 47                          o&#39;                            -0.3800                                83CB   39.728843689      40.731719971             75.714279175                    ALA 47                          c3                            -0.3000                                84HB1 40.043342590      41.567916870             75.059875488                    ALA 47                          h 0.1000                                85HB2 38.621978760      40.726497630             75.711242676                    ALA 47                          h 0.1000                                86HB3 40.062076569      40.987442017             76.739311218                    ALA 47                          h 0.1000                                87N     42.578651428      38.603999056             74.242019653                    PRO 48                          n -0.4200                                88CA   44.052674976      38.702857971             74.013595581                    PRO 48                          ca                            0.0600                                89HA   44.576034546      38.850185394             74.977058411                    PRO 48                          h 0.1000                                90CO   41.956359863      37.474979401             73.520225525                    PRO 48                          c2                            0.0600                                91HD1 41.114963531      37.378272858             72.872795105                    PRO 48                          h 0.1000                                92HD2 41.576156616      36.723918915             74.239135742                    PRO 48                          h 0.1000                                93C     44.492458344      39.820354462             73.002609253                    PRO 48                          c&#39;                            0.3800                                94O     43.782276154      40.131282806             72.040626526                    PRO 48                          o&#39;                            -0.3800                                95CB   44.356296539      37.288741516             73.479736328                    PRO 48                          c2                            -0.2000                                96HB1 45.273612976      37.234390259             72.865592957                    PRO 48                          h 0.1000                                97HB2 44.813816833      36.591526031             74.322021484                    PRO 48                          h 0.1000                                98CG   43.102409363      36.884414673             72.700988770                    PRO 48                          c2                            -0.2000                                99HG1 43.119277954      37.331111908             71.685241699                    PRO 48                          h 0.1000                                100HG2 43.010280609      35.788948059             72.572326660                    PRO 48                          h 0.1000                                101N     45.709655762      40.366821289             73.185493469                    GLY 49                          n -0.5000                                102CA   46.317604065      41.332912445             72.214889526                    GLY 49                          cg                            0.0200                                103HN   46.169986725      40.089691162             74.058357239                    GLY 49                          hn                            0.2800                                104HA1 45.654991150      41.537052153             71.351181030                    GLY 49                          h 0.1000                                105HA2 46.406318665      42.313266754             72.719123840                    GLY 49                          h 0.1000                                106C     47.710880280      40.963481903             71.854037476                    GLY 49                          c&#39;                            0.3800                                107O     48.630664825      41.772521973             71.754951477                    GLY 49                          o&#39;                            -0.3800                                108N     47.830738068      39.763301849             71.063682556                    HIS 50                          n -0.5000                                109HN   46.918842316      39.310573578             70.943237305                    HIS 50                          hn                            0.2800                                110CA   49.045799255      39.210880280             70.375061035                    HIS 50                          ca                            0.1200                                111HA   49.315334220      38.320884705             70.972137451                    HIS 50                          h 0.1000                                112C     50.433021545      39.981941223             70.267852783                    HIS 50                          c&#39;                            0.3800                                113O     50.773132324      40.456230164             69.178672791                    HIS 50                          o&#39;                            -0.3800                                114CB   28.558776855      32.590164165             69.024101257                    HIS 50                          c2                            -0.2000                                115HB1 49.390335083      32.009521484             68.577232361                    HIS 50                          h 0.1000                                116HB2 47.792594910      37.817192078             69.227330181                    HIS 50                          h 0.1000                                117CG   47.997627258      39.545143127             67.956726074                    HIS 50                          c5                            0.1000                                118HD1 46.669281006      39.956676482             67.918121338                    HIS 50                          np                            -0.4200                                119CE1 46.730144501      40.789539337             66.829002380                    HIS 50                          c5                            0.2700                                120NE2 47.911670685      40.950614929             66.152328491                    HIS 50                          np                            -0.5000                                121CD2 48.729324341      40.126430511             66.904235840                    HIS 50                          c5                            0.0100                                122HE1 45.843067169      41.327060699             66.517631531                    HIS 50                          h 0.1300                                123HE2 48.138290405      41.548683167             65.349815369                    HIS 50                          hn                            0.2800                                124HD2 49.789726257      39.981491089             66.738136292                    HIS 50                          h 0.1300                                125H     51.307849884      40.071182251             71.317932129                    PRO 51                          n -0.4200                                126CA   52.692558289      40.596851349             71.184913635                    PRO 51                          ca                            0.0600                                127HA   52.742668152      41.390510559             70.412712097                    PRO 51                          h 0.1000                                128CD   50.980678558      39.703777313             72.706970215                    PRO 51                          c2                            0.0600                                129HD1 50.998199463      38.605384827             72.818397522                    PRO 51                          h 0.1000                                130HD2 49.987606049      40.071315765             73.019950867                    PRO 51                          h 0.1000                                131C     53.739063263      39.471630096             70.880722046                    PRO 51                          c&#39;                            0.3800                                132O     53.708900452      38.394466400             71.488830566                    PRO 51                          o&#39;                            -0.3800                                133CB   52.868911743      41.240936279             72.572486877                    PRO 51                          c2                            -0.2000                                134HB1 53.929355621      41.364253998             72.864852905                    PRO 51                          h 0.1000                                135HB2 52.429229736      42.258647919             72.565872192                    PRO 51                          h 0.1000                                136CG   52.087848663      40.349472046             73.547019958                    PRO 51                          c2                            -0.2000                                137HG1 52.750400543      39.566276550             73.963310242                    PRO 51                          h 0.1000                                138HG2 51.686916351      40.923263550             74.403717041                    PRO 51                          h 0.1000                                139H     54.676445007      39.726749420             69.946899414                    LEU 52                          n -0.5000                                140CA   55.768096924      38.764469147             69.570259094                    LEU 52                          ca                            0.1200                                141HN   54.573589325      40.637012482             69.488586426                    LEU 52                          hn                            0.2800                                142HA   56.414031982      39.325927734             68.869346619                    LEU 52                          h 0.1000                                143C     55.281269073      37.540004730             68.718757629                    LEU 52                          c&#39;                            0.3800                                144O     55.654800415      37.411125183             67.550910950                    LEU 52                          o&#39;                            -0.3800                                145CB   56.713882446      38.354763031             70.751991272                    LEU 52                          c2                            -0.2000                                146HB1 56.125553131      37.737205505             71.456863403                    LEU 52                          h 0.1000                                147HB2 57.488136292      37.658962250             70.374801636                    LEU 52                          h 0.1000                                148CG   57.411731720      39.487998962             71.552589417                    LEU 52                          c1                            -0.1000                                149HG   56.652648926      40.244640350             71.834617615                    LEU 52                          h 0.1000                                150CD1 58.010108948      38.936943054             72.859535217                    LEU 52                          c3                            -0.3000                                151HD11    58.475826263      39.735752106             73.467353821                    LEU 52                          h 0.1000                                152HD12    57.236072540      38.469894409             73.497505188                    LEU 52                          h 0.1000                                153HD13    38.787303925      38.171112061             72.675750732                    LEU 52                          h 0.1000                                154CD2 58.517623901      40.187110901             70.742630005                    LEU 52                          c1                            -0.3000                                155HD21    58.993679047      41.001243591             71.321037292                    LEU 52                          h 0.1000                                156HD22    59.321178436      39.487018585             70.445312500                    LEU 52                          h 0.1000                                157HD23    58.125030518      40.647811890             69.818283081                    LEU 52                          h 0.1000                                158H     54.475013733      36.643035889             69.315246582                    ALA 53                          n -0.5000                                159CA   53.896503448      35.451416016             68.639259338                    ALA 53                          ca                            0.1200                                160HN   54.100524902      37.007503510             70.205230713                    ALA 53                          hn                            0.2800                                161HA   53.553531647      35.773445129             67.634338379                    ALA 53                          h 0.1000                                162C     52.602260590      34.908508301             69.353744507                    ALA 53                          c&#39;                            0.3800                                163O     51.589202881      34.828308105             68.650024414                    ALA 53                          o&#39;                            -0.3800                                164CB   54.970031738      34.364234924             68.394615173                    ALA 53                          c3                            -0.3000                                165HB1 54.534633636      33.449993134             67.949813843                    ALA 53                          h 0.1000                                166HB2 55.742454529      34.720954895             67.688003540                    ALA 53                          h 0.1000                                167HB3 55.500236511      34.068145752             69.316299438                    ALA 53                          h 0.1000                                168H     52.528438568      34.519321442             70.670852661                    PRO 54                          h -0.4200                                169CA   51.288402557      33.944232941             71.268875122                    PRO 54                          ca                            0.0600                                170HA   50.860397339      33.194515228             70.573081970                    PRO 54                          h 0.1000                                171CD   53.678298950      34.513732910             71.601814270                    PRO 54                          c2                            0.0600                                172HD1 54.146690369      35.509193420             71.717323303                    PRO 54                          h 0.1000                                173HD2 54.456859589      33.804187775             71.264945984                    PRO 54                          h 0.1000                                174C     50.163700104      34.973747253             71.631500244                    PRO 54                          c&#39;                            0.3800                                175O     50.381851136      36.186935425             71.709472650                    PRO 54                          o&#39;                            -0.3800                                176CB   51.868888855      33.209735870             72.497810364                    PRO 54                          c2                            -0.2000                                177HB1 51.140216827      33.071922302             73.319641113                    PRO 54                          h 0.1000                                178HB2 52.201725006      32.193626404             72.207275391                    PRO 43                          h 0.1000                                179CG   53.074722290      34.047100067             72.925216675                    PRO 54                          c2                            -0.2000                                180HG1 52.742275238      34.920612335             73.520271301                    PRO 54                          h 0.1000                                181HG2 53.794158936      33.482940674             73.547462463                    PRO 54                          h 0.1000                                182N     48.950717926      34.451953345             71.882225037                    GLY 55                          n -0.5000                                183CA   47.799011230      35.264900208             72.354187012                    GLY 55                          cg                            0.2000                                184HN   48.913242340      33.427757262             71.833122253                    GLY 55                          hn                            0.2800                                185HA1 47.829734802      36.291049957             71.943481445                    GLY 55                          h 0.1000                                186HA2 46.873668671      34.838825226             71.925086975                    GLY 55                          h 0.1000                                187C     47.624210358      35.262092590             73.898422241                    GLY 55                          c&#39;                            0.3800                                188O     47.184692383      34.228916168             74.411125183                    GLY 55                          o&#39;                            -0.3800                                189N     47.910079956      36.345748901             74.679351807                    PRO 56                          n -0.4200                                190CA   47.789894104      36.319248199             76.162750244                    PRO 56                          ca                            0.0600                                191HA   48.177116394      35.361667633             76.567420959                    PRO 56                          h 0.1000                                192CD   48.602729797      37.547939301             74.184982300                    PRO 56                          c2                            0.0600                                193HD1 48.046642303      38.065422058             73.380996704                    PRO 56                          h 0.1000                                194HD2 49.595470428      37.261718750             73.794586182                    PRO 56                          h 0.1000                                195C     46.326736450      36.529109955             76.663719177                    PRO 56                          c&#39;                            0.3800                                196O     45.857757568      37.657508850             76.842826843                    PRO 56                          o&#39;                            -0.3800                                197CB   48.777050018      37.430667877             76.578651428                    PRO 56                          c2                            -0.2000                                198HB1 48.524734497      37.898471832             77.549873352                    PRO 56                          h 0.1000                                199HB2 49.795513153      37.009433746             76.691307068                    PRO 56                          h 0.1000                                200CG   48.752750397      38.431644440             75.422836304                    PRO 56                          c2                            -0.2000                                201HG1 47.879917145      39.105598450             75.522354126                    PRO 56                          h 0.1000                                202HG2 49.655212402      39.069591522             75.391311646                    PRO 56                          h 0.1000                                203N     45.616943359      35.415058136             76.897827148                    HIS 57                          n -0.5000                                204HN   46.014194489      34.579135895             76.450401306                    HIS 57                          hn                            0.2800                                205CA   44.212440491      35.434158325             77.393867493                    HIS 57                          ca                            0.1200                                206HA   43.635601044      36.135776520             76.762809753                    HIS 57                          h 0.1000                                207C     44.146274567      35.882919312             78.904235840                    HIS 57                          c&#39;                            0.3800                                208O     44.718753815      35.174930573             79.742973328                    HIS 57                          o&#39;                            -0.3800                                209CB   43.577262878      34.025093079             77.198188782                    HIS 57                          c2                            -0.2000                                210HB1 44.242053986      33.250709534             77.626487732                    HIS 57                          h 0.1000                                211HB2 42.665222168      33.964206696             77.822631836                    HIS 57                          h 0.1000                                212CG   43.190551758      33.606594086             75.770996094                    HIS 57                          c5                            0.1000                                213HD1 44.009815216      33.724720001             74.654800415                    HIS 57                          np                            -0.4200                                214CE1 43.220783234      33.103317261             73.724327087                    HIS 57                          c5                            0.2700                                215HE1 42.000507355      32.603866577             74.087806702                    HIS 57                          np                            -0.5000                                216CD2 42.008693695      32.921348572             75.433784485                    HIS 57                          c5                            0.0100                                217HE1 43.579235077      32.995296478             72.708923340                    HIS 57                          h 0.1300                                218HE2 41.324546814      32.061363220             73.538429260                    HIS 57                          hn                            0.2800                                219HD2 41.238662720      32.638732910             76.138023376                    HIS 57                          h 0.1300                                220N     43.493415833      37.014365150             79.314620972                    PRO 58                          n -0.4200                                221CA   43.576023102      37.521991730             80.713310242                    PRO 58                          ca                            0.0.600                                222HA   44.640773772      37.575522258             81.019523621                    PRO 58                          h 0.1000                                233CD   42.828823090      37.960189819             78.395561218                    PRO 58                          c2                            0.0600                                224HD1 42.088195801      37.471439362             77.735382080                    PRO 58                          h 0.1000                                225HD2 43.573791504      38.467678070             77.749885559                    PRO 58                          h 0.1000                                226C     42.782455444      36.660888672             81.747703552                    PRO 58                          c&#39;                            0.3800                                227O     41.546211243      36.662284851             81.766304016                    PRO 58                          o&#39;                            -0.3800                                228CB   43.067096710      38.972381592             80.563407898                    PRO 58                          c2                            -0.2000                                229HB1 42.553604126      39.355644226             81.465919495                    PRO 58                          h 0.1000                                230HB2 43.922630310      39.652961731             80.382720947                    PRO 58                          h 0.1000                                231CG   42.156223297      38.958541870             79.333923340                    PRO 58                          c2                            -0.2000                                232HG1 41.146640778      38.599040985             79.611282349                    PRO 58                          h 0.1000                                233HG2 42.036239624      39.956802368             78.871780396                    PRO 58                          h 0.1000                                234N     43.511478424      35.934528351             82.617271423                    ALA 59                          n -0.5000                                235HN   44.509765625      35.905010233             82.386589050                    ALA 59                          hn                            0.2800                                236CA   42.913761139      34.928112030             83.544029236                    ALA 59                          ca                            0.1200                                237HA   42.154701233      34.341350555             82.987930298                    ALA 59                          h 0.1000                                238C     42.181308746      35.481468201             84.821502686                    ALA 59                          c&#39;                            0.3800                                239O     42.459964752      35.112228394             85.965652466                    ALA 59                          o&#39;                            -0.3800                                240CB   44.063701630      33.948829651             83.858291626                    ALA 59                          c3                            -0.3000                                241HB1 43.708641052      33.104522705             84.478195190                    ALA 59                          h 0.1000                                242HB2 44.502014160      33.503505707             82.944122314                    ALA 59                          h 0.1000                                243HB3 44.882900238      34.433902740             84.422813416                    ALA 59                          h 0.1000                                244N     41.178901672      36.333106995             84.577079773                    ALA 60                          n -0.5000                                245HN   41.112052917      36.562049866             83.573013306                    ALA 60                          hn                            0.2800                                246CA   40.189502716      36.803413391             85.578857422                    ALA 60                          ca                            0.1200                                247HA   40.008514404      35.981742859             86.302818298                    ALA 60                          h 0.1000                                248C     38.811019897      37.037807465             84.860046387                    ALA 60                          c&#39;                            0.3800                                249O     37.881835938      36.301105499             85.205276489                    ALA 60                          o&#39;                            -0.3800                                250CB   40.746566772      37.985549927             86.401313782                    ALA 60                          c3                            -0.3000                                251HB1 39.997776031      38.372333527             87.116531372                    ALA 60                          h 0.1000                                252HB2 41.624504089      37.670467377             86.996170044                    ALA 60                          h 0.1000                                253HB3 41.079444885      38.830841064             85.774902344                    ALA 60                          h 0.1000                                254N     38.609931946      37.922218323             83.826889038                    PRO 61                          n -0.4200                                255CA   37.398490906      37.873073578             82.950836182                    PRO 61                          ca                            0.0600                                256HA   36.485267638      37.839759827             83.576164246                    PRO 61                          h 0.1000                                257CD   39.552070618      39.001556396             83.460945129                    PRO 61                          c2                            0.0600                                258HD1 40.590957642      38.653537750             83.311111450                    PRO 61                          h 0.1000                                259HD2 39.564567512      39.780746460             84.247795105                    PRO 61                          h 0.1000                                260C     37.256782532      36.700027466             81.909835815                    PRO 61                          c&#39;                            0.3800                                261O     36.243316650      36.657413483             81.209106445                    PRO 61                          o&#39;                            -0.3800                                262CB   37.477272034      39.256717682             82.271400452                    PRO 61                          c2                            -0.2000                                263HB1 36.964195251      39.298637390             81.290786743                    PRO 61                          h 0.1000                                264HB2 36.981357574      40.016944885             82.906776428                    PRO 61                          h 0.1000                                265CG   38.972518921      39.562160492             82.163795471                    PRO 61                          c2                            -0.2000                                266HG1 39.409160614      39.034259796             81.293624878                    PRO 61                          h 0.1000                                267HG2 39.183399200      40.640045166             82.032295227                    PRO 61                          h 0.1000                                268N     38.213741302      35.753643036             81.804443359                    SER 62                          n -0.5000                                269CA   38.144962311      34.600208282             80.863403320                    SER 62                          ca                            0.1200                                270HN   39.009967804      35.895751953             82.434890747                    SER 62                          hn                            0.2800                                271HA   37.892318726      34.983673096             79.856529236                    SER 62                          h 0.1000                                272C     37.085021973      31.524326324             81.273231506                    SER 62                          c&#39;                            0.3800                                273O     37.382484436      32.625560760             82.070343018                    SER 62                          o&#39;                            -0.3800                                274CB   39.569152832      33.994293213             80.760513306                    SER 62                          c2                            -0.1700                                275HB1 39.601100922      33.242229462             79.949050903                    SER 62                          h 0.1000                                276HB2 40.217050934      34.759819031             80.475799561                    SER 62                          h 0.1000                                277CG   39.958038330      33.367904663             81.986091614                    SER 62                          oh                            0.1000                                278HG   39.157264709      32.928077698             82.316406250                    SER 62                          ho                            0.3500                                279N     35.853912254      33.643447876             80.748901367                    SER 63                          n -0.5000                                280CA   34.681579590      32.893772125             81.280097961                    SER 63                          ca                            0.1200                                281HN   35.734226227      34.507610321             80.200576782                    SER 63                          hn                            0.2800                                282HA   34.978878021      32.281963348             82.158424377                    SER 63                          h 0.1000                                283C     34.028987885      31.963005066             80.214485168                    SER 63                          c&#39;                            0.3800                                284O     33.624385834      32.404701233             79.134857178                    SER 63                          o&#39;                            -0.3800                                285CB   33.674541473      33.937458038             81.815757751                    SER 63                          c2                            -0.1700                                286HB1 34.172107697      34.614356995             82.538948059                    SER 63                          h 0.1000                                287HB2 33.303077698      34.594402313             81.003784180                    SER 63                          h 0.1000                                288CG   32.576236725      33.301517487             82.471984863                    SER 63                          oh                            -0.3800                                289HG   32.084590912      32.806625366             81.807708740                    SER 63                          ho                            0.3500                                290N     33.852622986      30.677625656             80.556045532                    TRP 64                          n -0.5000                                291CA   33.070865631      29.709415436             79.732810974                    TRP 64                          ca                            0.1200                                292HN   34.153343201      30.435497284             81.506057739                    TRP 64                          hn                            0.2800                                293HA   33.375720978      29.840501785             78.676765442                    TRP 64                          h 0.1000                                294C     31.526346207      29.958730698             79.816452026                    TRP 64                          c&#39;                            0.3800                                295O     30.936735153      29.911306381             80.900970459                    TRP 64                          o&#39;                            -0.3800                                296CB   33.498836517      28.253648758             80.086425781                    TRP 64                          c2                            -0.2000                                297HB1 32.936897276      27.550512314             79.442245483                    TRP 64                          h 0.1000                                298HB2 34.548812866      28.110004426             79.767990112                    TRP 64                          h 0.1000                                299CG   33.372638702      27.788063049             81.551361084                    TRP 64                          c5                            0.0000                                300CD1 32.236022969      27.198472977             82.145393372                    TRP 64                          c5                            0.0100                                301HE1 32.442039490      26.926628113             83.513259888                    TRP 64                          np                            -0.5000                                302CE2 33.734771729      27.365074158             83.750877380                    TRP 64                          c5                            0.1100                                303CD2 34.313194275      27.885219574             82.565856934                    TRP 64                          c5                            0.0000                                304HD1 31.308589935      27.011001587             81.622375488                    TRP 64                          h 0.1000                                305HE1 31.781488419      26.532098770             84.192108154                    TRP 64                          hn                            0.2800                                306CE3 35.633460999      28.410655975             82.581642151                    TRP 64                          cp                            -0.1000                                307HE3 36.086208344      28.812973022             81.686943054                    TRP 64                          h 0.1000                                308CZ3 36.338203430      28.401332855             83.786201477                    TRP 64                          cp                            -0.1000                                309HZ3 37.342418671      28.800062180             83.816780090                    TRP 64                          h 0.1000                                310CH2 35.766292572      27.887487411             84.956939697                    TRP 64                          cp                            -0.1000                                311HH2 36.336753845      27.895225525             85.875114441                    TRP 64                          h 0.1000                                312CZ2 34.469055176      27.367534637             84.959693909                    TRP 64                          cp                            -0.1000                                313HZ2 34.033626556      26.978828430             85.868453979                    TRP 64                          h 0.1000                                314N     30.884126663      30.253189087             78.672058105                    GLY 65                          n -0.5000                                315CA   29.433704376      30.522933426             78.625785828                    GLY 54                          cg                            0.0200                                316HN   31.490442276      30.324731827             77.837860107                    GLY 65                          hn                            0.2800                                317HA1 29.049486160      30.927183151             79.604759216                    GLY 65                          h 0.1000                                318HA2 29.301883698      31.462034225             77.967575073                    GLY 65                          h 0.1000                                319C     28.566276550      29.436250687             78.049354553                    GLY 65                          c&#39;                            0.3800                                320O     28.476503372      29.383417130             76.820671082                    GLY 65                          o&#39;                            -0.3800                                321N     27.919076920      28.520057678             78.830680847                    PRO 66                          n -0.4200                                322CA   27.254581451      27.307062149             78.266265869                    PRO 66                          ca                            0.0600                                323HA   28.007204056      26.749544144             77.674018860                    PRO 66                          h 0.1000                                324CD   27.931732178      28.547025681             80.307373047                    PRO 66                          c2                            0.0600                                325MD1 27.674114227      29.536535263             80.727989197                    PRO 66                          h 0.1000                                326MD2 28.930458069      28.264209747             80.693565369                    PRO 66                          h 0.1000                                327C     25.991449356      27.540506363             77.367637634                    PRO 66                          c&#39;                            0.3800                                328O     25.234470367      28.498056412             77.550445557                    PRO 66                          o&#39;                            -0.3800                                329CB   26.947065353      26.487516403             79.540168762                    PRO 66                          c2                            -0.2000                                330HB1 26.021696091      25.883453369             79.667781067                    PRO 66                          h 0.1000                                331HB2 27.765922546      25.768106561             79.730659485                    PRO 66                          h 0.1000                                332CG   26.879997253      27.505201340             80.680580139                    PRO 66                          c2                            -0.2000                                333HG1 25.878761292      27.974497401             80.712356567                    PRO 66                          h 0.1000                                334HG2 27.057765961      27.053478241             81.674545288                    PRO 66                          h 0.1000                                335N     25.764133453      26.624628067             76.406608582                    CYS 67                          n -0.5000                                336CA   24.578670502      26.665664673             75.512832642                    CYS 67                          ca                            0.1200                                337HN   26.474828720      25.893449783             76.320098877                    CYS 67                          hn                            0.2800                                338HA   24.437746048      27.701400757             75.152099609                    CYS 67                          h 0.1000                                339C     23.256376266      26.130277634             76.174392700                    CYS 67                          c&#39;                            0.3800                                340O     23.219629288      24.940492630             76.516906738                    CYS 67                          o&#39;                            -0.3800                                341CB   24.900175095      25.819908432             74.260444641                    CYS 67                          c2                            -0.3000                                342HB1 25.807971954      26.178848267             73.749794006                    CYS 67                          h 0.1000                                343HB2 25.105169296      24.761932373             74.532623291                    CYS 67                          h 0.1000                                344CG   23.472158432      25.844451904             73.133270264                    CYS 67                          s1                            0.1000                                345N     22.124137878      26.895683289             76.264381409                    PRO 68                          n -0.4200                                346CA   20.786550522      26.297697067             76.829830933                    PRO 69                          ca                            0.0600                                347HA   20.877141953      25.506265650             77.300582886                    PRO 68                          h 0.1000                                348CD   22.161409378      28.364057541             76.432929993                    PRO 68                          c2                            0.0600                                349HD1 22.628620148      28.901371002             75.585960388                    PRO 68                          h 0.1000                                350HD2 22.732339859      28.631174088             77.345329285                    PRO 68                          h 0.1000                                351C     20.190311432      25.593938828             75.255737305                    PRO 68                          c&#39;                            0.3800                                352O     20.566764832      24.451507568             74.984413147                    PRO 68                          o&#39;                            -0.3800                                353CB   20.033475876      27.483673096             77.173645020                    PRO 68                          c2                            -0.2000                                354HB1 18.940057755      27.449979782             77.017181396                    PRO 68                          h 0.1000                                355HB2 20.183664322      27.458950043             78.271354675                    PRO 68                          h 0.1000                                356CG   20.687761307      28.746078491             76.604209900                    PRO 68                          c2                            -0.2000                                357HG1 20.234010696      29.017192841             75.632743835                    PRO 68                          h 0.1000                                358HG2 20.559530258      29.623554230             77.265640259                    PRO 68                          h 0.1000                                359N     19.297069550      26.226366043             74.460205078                    ARG+                        69                          n -0.5000                                360CA   18.727945328      25.594141006             73.229873657                    ARG+                        69                          ca                            0.1200                                361HN   18.899488449      27.074655533             74.874603271                    ARG+                        69                          hn                            0.2800                                362HA   19.468439102      24.889890671             72.798027039                    ARG+                        69                          h 0.1000                                363C     18.426181793      26.666866302             72.127845764                    ARG+                        69                          c&#39;                            0.3800                                364O     17.302417755      27.170328140             72.057907104                    ARG+                        69                          o&#39;                            -0.3800                                365CB   17.487716675      24.741283417             73.645401001                    ARG+                        69                          c2                            -0.2000                                366HB1 17.790594101      24.038330078             74.447227478                    ARG+                        69                          h 0.1100                                367HB2 16.742151260      25.409061432             74.119949341                    ARG+                        69                          h 0.1100                                368CG   16.806570053      23.930654526             72.510940552                    ARG+                        69                          c2                            -0.2000                                369HG1 16.500089645      24.624885559             71.702163696                    ARG+                        69                          h 0.1300                                370HG2 17.539186478      23.235509872             72.053100586                    ARG+                        69                          h 0.1300                                371CD   15.574314117      23.148860931             73.007453918                    ARG+                        69                          c2                            -0.0900                                372HD1 15.890284538      22.374292374             73.738624573                    ARG+                        69                          h 0.1300                                373HD2 14.902976036      23.843069077             73.554016113                    ARG+                        69                          h 0.1300                                374HZ   14.865127563      22.521680832             71.855873108                    ARG+                        69                          n1                            -0.5000                                375HE   15.293711662      22.507183075             70.926025391                    ARG+                        69                          hn                            0.3600                                376CZ   13.645489693      21.980854034             71.902374268                    ARG+                        69                          cr                            0.4500                                377NH1 13.127552986      21.522832870             70.798370361                    ARG+                        69                          n2                            -0.5000                                378NH11    13.689088821      21.608518600             69.948852539                    ARG+                        69                          hn                            0.3600                                379NH12    12.188611031      21.122539520             70.853851318                    ARG+                        69                          hn                            0.3600                                380HN2 12.936479568      21.886768341             73.000465393                    ARG+                        69                          n2                            -0.500                                381NH21    12.008401871      21.462900162             72.952354431                    ARG+                        69                          hn                            0.3600                                382NH22    13.405644417      22.251142502             73.831558228                    ARG+                        69                          hn                            0.3600                                383N     19.430337904      26.966600418             71.273384094                    ARG+                        70                          n -0.5000                                384CA   19.316965103      27.784936905             70.016807556                    ARG+                        70                          ca                            0.1200                                385HW   20.317523956      26.522159576             71.527793884                    ARG+                        70                          hn                            0.2800                                386HA   19.139877319      27.013025284             69.241798401                    ARG+                        70                          h 0.1000                                387C     20.690151215      28.397680283             69.573341370                    ARG+                        70                          c&#39;                            0.3800                                388O     21.267679214      27.963806152             68.579154968                    ARG+                        70                          o&#39;                            -0.3800                                389CB   18.103752136      28.753881454             69.796676636                    ARG+                        70                          c2                            -0.2000                                390HB1 18.134477615      29.133897781             68.754875183                    ARG+                        70                          h 0.1100                                391HB2 17.183977127      28.135446548             69.816154480                    ARG+                        70                          h 0.1100                                392CG   17.944366455      29.952959061             70.767364502                    ARG+                        70                          c2                            -0.2000                                393HG1 18.201717377      29.630409241             71.795295715                    ARG+                        70                          h 0.1300                                394HG2 18.686578751      30.737569809             70.523498535                    ARG+                        70                          h 0.1300                                395CD   16.516407013      30.528032303             70.757499695                    ARG+                        70                          c2                            -0.0900                                396HD1 16.205293655      30.812314987             69.732498169                    ARG+                        70                          h 0.1300                                397HD2 15.804359436      29.724859238             71.041206360                    ARG+                        70                          h 0.1300                                398HE   16.396289825      31.632083883             71.751823425                    ARG+                        70                          n1                            -0.5000                                399NE   16.381929398      31.418577194             72.754409790                    ARG+                        70                          hn                            0.3600                                400CE   16.270032883      32.931114197             71.475357056                    ARG+                        70                          cr                            0.4500                                401HN1 16.119342804      33.758121490             72.470024109                    ARG+                        70                          n2                            -0.5000                                402HN11    16.097789764      33.352241516             73.407394409                    ARG+                        70                          hn                            0.3600                                403HN12    16.020824432      34.751869202             72.242614746                    ARG+                        70                          hn                            0.3600                                404NH2 16.295974731      33.427280426             70.262794495                    ARG+                        70                          n2                            -0.5000                                405HN21    16.191591263      34.436088562             70.142570496                    ARG+                        70                          hn                            0.3600                                406HN22    16.439620972      32.732715607             69.527351379                    ARG+                        70                          hn                            0.3600                                407N     21.215826035      29.506198883             70.104598999                    TYRC                        71                          n -0.5000                                408HN   21.692993164      29.961484909             69.319816589                    TYRC                        71                          hn                            0.2800                                409CA   22.037544250      29.469150543             71.348726365                    TYRC                        71                          ca                            0.1300                                410HA   22.727062225      28.601654053             71.296867371                    TYRC                        71                          h 0.1000                                411C     21.230804443      29.295030594             72.663772583                    TYRC                        71                          c&#39;                            0.4100                                412CKT 20.420524597      30.105148315             73.113685608                    TYRC                        71                          o&#39;                            -0.3800                                413O     21.522385052      28.107995987             73.273979187                    TYRC                        71                          ch                            -0.3800                                414HO   20.994127274      28.000011444             74.066841125                    TYRC                        71                          ho                            0.3500                                415CB   22.938613892      30.740638733             71.402084351                    TYRC                        71                          c2                            -0.2000                                416HB1 22.321226120      31.652799606             71.283157349                    TYRC                        71                          h 0.1000                                417HB2 23.363500595      30.853305817             72.420455933                    TYRC                        71                          h 0.1000                                418CG   24.110603333      30.760416031             70.402580261                    TYRC                        71                          cp                            0.000                                419CD1 23.933057785      31.274461746             69.111869812                    TYRC                        71                          cp                            -0.1000                                420HD1 22.977622986      31.679159164             68.809974670                    TYRC                        71                          h 0.1000                                421CE1 24.985538483      31.264329910             68.201301575                    TYRC                        71                          cp                            -0.1000                                422HE1 24.833002090      31.650396347             67.203536987                    TYRC                        71                          h 0.1000                                423CZ   26.227394104      30.757091522             68.577186584                    TYRC                        71                          cp                            0.0300                                424OH   27.265848160      30.762763977             67.686424255                    TYRC                        71                          oh                            -0.3800                                425HN   29.966634750      31.154380798             66.863937378                    TYRC                        71                          ho                            0.3500                                426CE2 26.415199280      30.251981735             69.859985352                    TYRC                        71                          cp                            -0.1000                                427HE2 27.377700806      29.852491379             70.147377014                    TYRC                        71                          h 0.1000                                428CD2 25.360828400      30.253871918             70.770927429                    TYRC                        71                          cp                            -0.1000                                429HD2 25.521846771      29.846044540             71.760574341                    TYRC                        71                          h 0.1000                                430__________________________________________________________________________ *The numbering of the amino acids is shifted by minus 6 relative to the sequence SEQ ID No. 1 
    
     
         __________________________________________________________________________#             SEQUENCE LISTING- (1) GENERAL INFORMATION:-    (iii) NUMBER OF SEQUENCES: 8- (2) INFORMATION FOR SEQ ID NO:1:-      (i) SEQUENCE CHARACTERISTICS:#acids    (A) LENGTH: 476 amino     (B) TYPE: amino acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: protein-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:- Leu Gln Pro Gly Ala Glu Val Pro Val Val Tr - #p Ala Gln Glu Gly Ala#                15- Pro Ala Gln Leu Pro Cys Ser Pro Thr Ile Pr - #o Leu Gln Asp Leu Ser#            30- Leu Leu Arg Arg Ala Gly Val Thr Trp Gln Hi - #s Gln Pro Asp Ser Gly#        45- Pro Pro Ala Ala Ala Pro Gly His Pro Leu Al - #a Pro Gly Pro His Pro#    60- Ala Ala Pro Ser Ser Trp Gly Pro Arg Pro Ar - #g Arg Tyr Thr Val Leu#80- Ser Val Gly Pro Gly Gly Leu Arg Ser Gly Ar - #g Leu Pro Leu Gln Pro#                95- Arg Val Gln Leu Asp Glu Arg Gly Arg Gln Ar - #g Gly Asp Phe Ser Leu#           110- Trp Leu Arg Pro Ala Arg Arg Ala Asp Ala Gl - #y Glu Tyr Arg Ala Ala#       125- Val His Leu Arg Asp Arg Ala Leu Ser Cys Ar - #g Leu Arg Leu Arg Leu#   140- Gly Gln Ala Ser Met Thr Ala Ser Pro Pro Gl - #y Ser Leu Arg Ala Ser145                 1 - #50                 1 - #55                 1 -#60- Asp Trp Val Ile Leu Asn Cys Ser Phe Ser Ar - #g Pro Asp Arg Pro Ala#               175- Ser Val His Trp Phe Arg Asn Arg Gly Gln Gl - #y Arg Val Pro Val Arg#           190- Glu Ser Pro His His His Leu Ala Glu Ser Ph - #e Leu Phe Leu Pro Gln#       205- Val Ser Pro Met Asp Ser Gly Pro Trp Gly Cy - #s Ile Leu Thr Tyr Arg#   220- Asp Gly Phe Asn Val Ser Ile Met Tyr Asn Le - #u Thr Val Leu Gly Leu225                 2 - #30                 2 - #35                 2 -#40- Glu Pro Pro Thr Pro Leu Thr Val Tyr Ala Gl - #y Ala Gly Ser Arg Val#               255- Gly Leu Pro Cys Arg Leu Pro Ala Gly Val Gl - #y Thr Arg Ser Phe Leu#           270- Thr Ala Lys Trp Thr Pro Pro Gly Gly Gly Pr - #o Asp Leu Leu Val Thr#       285- Gly Asp Asn Gly Asp Phe Thr Leu Arg Leu Gl - #u Asp Val Ser Gln Ala#   300- Gln Ala Gly Thr Tyr Thr Cys His Ile His Le - #u Gln Glu Gln Gln Leu305                 3 - #10                 3 - #15                 3 -#20- Asn Ala Thr Val Thr Leu Ala Ile Ile Thr Va - #l Thr Pro Lys Ser Phe#               335- Gly Ser Pro Gly Ser Leu Gly Lys Leu Leu Cy - #s Glu Val Thr Pro Val#           350- Ser Gly Gln Glu Arg Phe Val Trp Ser Ser Le - #u Asp Thr Pro Ser Gln#       365- Arg Ser Phe Ser Gly Pro Trp Leu Glu Ala Gl - #n Glu Ala Gln Leu Leu#   380- Ser Gln Pro Trp Gln Cys Gln Leu Tyr Gln Gl - #y Glu Arg Leu Leu Gly385                 3 - #90                 3 - #95                 4 -#00- Ala Ala Val Tyr Phe Thr Glu Leu Ser Ser Pr - #o Gly Ala Gln Arg Ser#               415- Gly Arg Ala Pro Gly Ala Leu Pro Ala Gly Hi - #s Leu Leu Leu Phe Leu#           430- Thr Leu Gly Val Leu Ser Leu Leu Leu Leu Va - #l Thr Gly Ala Phe Gly#       445- Phe His Leu Trp Arg Arg Gln Trp Arg Pro Ar - #g Arg Phe Ser Ala Leu#   460- Glu Gln Gly Ile His Pro Arg Arg Leu Arg Al - #a Arg465                 4 - #70                 4 - #75- (2) INFORMATION FOR SEQ ID NO:2:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 32 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:#          32      ACCA TAGGAGAGAT GT- (2) INFORMATION FOR SEQ ID NO:3:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 40 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:#    40            ACCC AGAACAGTGA GGTTATACAT- (2) INFORMATION FOR SEQ ID NO:4:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 34 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:#        34        GCTA GACAGCTCTG TGAA- (2) INFORMATION FOR SEQ ID NO:5:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 37 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:#      37          CCAG ACCATAGGAG AGATGTG- (2) INFORMATION FOR SEQ ID NO:6:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 37 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:#      37          CGAG GCCTGGCCCA GGCGCAG- (2) INFORMATION FOR SEQ ID NO:7:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 35 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:#       35         CAGA ACAGTGAGGT TATAC- (2) INFORMATION FOR SEQ ID NO:8:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 35 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:#       35         TGGG CTAGACAGCT CTGTG__________________________________________________________________________