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HEADER    COMPLEX (LYASE/PEPTIDE)                 15-MAY-96   1YAS
TITLE     HYDROXYNITRILE LYASE COMPLEXED WITH HISTIDINE
COMPND    MOL_ID: 1;
COMPND   2 MOLECULE: HYDROXYNITRILE LYASE;
COMPND   3 CHAIN: A;
COMPND   4 SYNONYM: OXYNITRILE LYASE;
COMPND   5 EC: 4.2.1.39;
COMPND   6 ENGINEERED: YES;
COMPND   7 MOL_ID: 2;
COMPND   8 MOLECULE: HISTIDINE;
COMPND   9 CHAIN: B
COMPND  10 ENGINEERED: YES
SOURCE    MOL_ID: 1;
SOURCE   2 ORGANISM_SCIENTIFIC: HEVEA BRASILIENSIS;
SOURCE   3 ORGANISM_COMMON: RUBBER TREE;
SOURCE   4 ORGAN: LEAF;
SOURCE   5 EXPRESSION_SYSTEM: PICHIA PASTORIS;
SOURCE   6 EXPRESSION_SYSTEM_PLASMID: BHIL-D2;
SOURCE   7 EXPRESSION_SYSTEM_GENE: HNL;
SOURCE   8 MOL_ID: 2;
SOURCE   9 SYNTHETIC: YES
KEYWDS    OXYNITRILASE, CYANOGENESIS, CYANHYDRIN FORMATION, LYASE,
KEYWDS   2 COMPLEX (LYASE/PEPTIDE)
EXPDTA    X-RAY DIFFRACTION
AUTHOR    U.G.WAGNER,C.KRATKY
REVDAT   1   16-JUN-97 1YAS    0
JRNL        AUTH   U.G.WAGNER,M.HASSLACHER,H.GRIENGL,H.SCHWAB,C.KRATKY
JRNL        TITL   MECHANISM OF CYANOGENESIS: THE CRYSTAL STRUCTURE OF
JRNL        TITL 2 HYDROXYNITRILE LYASE FROM HEVEA BRASILIENSIS
JRNL        REF    STRUCTURE (LONDON)            V.   4   811 1996
JRNL        REFN   ASTM STRUE6  UK ISSN 0969-2126                 2005
REMARK   1
REMARK   1 REFERENCE 1
REMARK   1  AUTH   M.HASSLACHER,C.KRATKY,H.GRIENGL,H.SCHWAB,
REMARK   1  AUTH 2 S.D.KOHLWEIN
REMARK   1  TITL   HYDROXYNITRILE LYASE FROM HEVEA BRASILIENSIS:
REMARK   1  TITL 2 MOLECULAR CHARACTERIZATION AND MECHANISM OF ENZYME
REMARK   1  TITL 3 CATALYSIS
REMARK   1  REF    PROTEINS: STRUCT.,FUNCT.,     V.  27   438 1997
REMARK   1  REF  2 GENET.
REMARK   1  REFN   ASTM PSFGEY  US ISSN 0887-3585                 0867
REMARK   1 REFERENCE 2
REMARK   1  AUTH   U.G.WAGNER,M.SCHALL,M.HAYN,M.HASSLACHER,H.SCHWAB,
REMARK   1  AUTH 2 H.S.GRIENGL,C.KRATKY
REMARK   1  TITL   CRYSTALLIZATION OF A HYDROXYNITRILE LYASE
REMARK   1  REF    ACTA CRYSTALLOGR.,SECT.D      V.  52   591 1996
REMARK   1  REFN   ASTM ABCRE6  DK ISSN 0907-4449                 0766
REMARK   1 REFERENCE 3
REMARK   1  AUTH   M.HASSLACHER,M.SCHALL,M.HAYN,H.GRIENGL,
REMARK   1  AUTH 2 S.D.KOHLWEIN,H.SCHWAB
REMARK   1  TITL   MOLECULAR CLONING OF THE FULL-LENGTH CDNA OF
REMARK   1  TITL 2 (S)-HYDROXYNITRILE LYASE FROM HEVEA BRASILIENSIS.
REMARK   1  TITL 3 FUNCTIONAL EXPRESSION IN ESCHERICHIA COLI AND
REMARK   1  TITL 4 SACCHAROMYCES CEREVISIAE AND IDENTIFICATION OF AN
REMARK   1  TITL 5 ACTIVE SITE RESIDUE
REMARK   1  REF    J.BIOL.CHEM.                  V. 271  5884 1996
REMARK   1  REFN   ASTM JBCHA3  US ISSN 0021-9258                 0071
REMARK   2
REMARK   2 RESOLUTION. 1.9  ANGSTROMS.
REMARK   3
REMARK   3 REFINEMENT.
REMARK   3   PROGRAM     : X-PLOR 3.1
REMARK   3   AUTHORS     : BRUNGER
REMARK   3
REMARK   3  DATA USED IN REFINEMENT.
REMARK   3   RESOLUTION RANGE HIGH (ANGSTROMS) : 1.9
REMARK   3   RESOLUTION RANGE LOW  (ANGSTROMS) : 15.
REMARK   3   DATA CUTOFF            (SIGMA(F)) : 3.0
REMARK   3   DATA CUTOFF HIGH         (ABS(F)) : NULL
REMARK   3   DATA CUTOFF LOW          (ABS(F)) : NULL
REMARK   3   COMPLETENESS (WORKING+TEST)   (%) : 76.7
REMARK   3   NUMBER OF REFLECTIONS             : 20547
REMARK   3
REMARK   3  FIT TO DATA USED IN REFINEMENT.
REMARK   3   CROSS-VALIDATION METHOD          : FREE-R
REMARK   3   FREE R VALUE TEST SET SELECTION  : NULL
REMARK   3   R VALUE            (WORKING SET) : 0.199
REMARK   3   FREE R VALUE                     : 0.241
REMARK   3   FREE R VALUE TEST SET SIZE   (%) : 10.
REMARK   3   FREE R VALUE TEST SET COUNT      : 2030
REMARK   3   ESTIMATED ERROR OF FREE R VALUE  : NULL
REMARK   3
REMARK   3  FIT IN THE HIGHEST RESOLUTION BIN.
REMARK   3   TOTAL NUMBER OF BINS USED           : NULL
REMARK   3   BIN RESOLUTION RANGE HIGH       (A) : 1.9
REMARK   3   BIN RESOLUTION RANGE LOW        (A) : 1.99
REMARK   3   BIN COMPLETENESS (WORKING+TEST) (%) : 76.68
REMARK   3   REFLECTIONS IN BIN    (WORKING SET) : 1230
REMARK   3   BIN R VALUE           (WORKING SET) : 0.338
REMARK   3   BIN FREE R VALUE                    : 0.381
REMARK   3   BIN FREE R VALUE TEST SET SIZE  (%) : 7.58
REMARK   3   BIN FREE R VALUE TEST SET COUNT     : 139
REMARK   3   ESTIMATED ERROR OF BIN FREE R VALUE : NULL
REMARK   3
REMARK   3  NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
REMARK   3   PROTEIN ATOMS            : 2057
REMARK   3   NUCLEIC ACID ATOMS       : 0
REMARK   3   HETEROGEN ATOMS          : 16
REMARK   3   SOLVENT ATOMS            : 77
REMARK   3
REMARK   3  B VALUES.
REMARK   3   FROM WILSON PLOT           (A**2) : 15.296
REMARK   3   MEAN B VALUE      (OVERALL, A**2) : 8.75
REMARK   3   OVERALL ANISOTROPIC B VALUE.
REMARK   3    B11 (A**2) : -2.856
REMARK   3    B22 (A**2) : 1.298
REMARK   3    B33 (A**2) : 1.558
REMARK   3    B12 (A**2) : 0.000
REMARK   3    B13 (A**2) : 0.000
REMARK   3    B23 (A**2) : 0.000
REMARK   3
REMARK   3  ESTIMATED COORDINATE ERROR.
REMARK   3   ESD FROM LUZZATI PLOT        (A) : NULL
REMARK   3   ESD FROM SIGMAA              (A) : NULL
REMARK   3   LOW RESOLUTION CUTOFF        (A) : 15.
REMARK   3
REMARK   3  CROSS-VALIDATED ESTIMATED COORDINATE ERROR.
REMARK   3   ESD FROM C-V LUZZATI PLOT    (A) : NULL
REMARK   3   ESD FROM C-V SIGMAA          (A) : NULL
REMARK   3
REMARK   3  RMS DEVIATIONS FROM IDEAL VALUES.
REMARK   3   BOND LENGTHS                 (A) : 0.018
REMARK   3   BOND ANGLES            (DEGREES) : 2.990
REMARK   3   DIHEDRAL ANGLES        (DEGREES) : 24.471
REMARK   3   IMPROPER ANGLES        (DEGREES) : 1.388
REMARK   3
REMARK   3  ISOTROPIC THERMAL MODEL : NULL
REMARK   3
REMARK   3  ISOTROPIC THERMAL FACTOR RESTRAINTS.    RMS    SIGMA
REMARK   3   MAIN-CHAIN BOND              (A**2) : NULL  ; NULL
REMARK   3   MAIN-CHAIN ANGLE             (A**2) : NULL  ; NULL
REMARK   3   SIDE-CHAIN BOND              (A**2) : NULL  ; NULL
REMARK   3   SIDE-CHAIN ANGLE             (A**2) : NULL  ; NULL
REMARK   3
REMARK   3  NCS MODEL : NULL
REMARK   3
REMARK   3  NCS RESTRAINTS.                         RMS   SIGMA/WEIGHT
REMARK   3   GROUP  1  POSITIONAL            (A) : NULL  ; NULL
REMARK   3   GROUP  1  B-FACTOR           (A**2) : NULL  ; NULL
REMARK   3
REMARK   3  PARAMETER FILE  1  : PARAM19X.PRO
REMARK   3  PARAMETER FILE  2  : PARAM19X.SOL
REMARK   3  TOPOLOGY FILE  1   : TOPH19X.PRO
REMARK   3  TOPOLOGY FILE  2   : TOPH19X.SOL
REMARK   3
REMARK   3  OTHER REFINEMENT REMARKS: NULL
REMARK   4
REMARK   4 1YAS COMPLIES WITH FORMAT V. 2.2, 16-DEC-1996
REMARK 200
REMARK 200 EXPERIMENTAL DETAILS
REMARK 200  EXPERIMENT TYPE                : X-RAY DIFFRACTION
REMARK 200  DATE OF DATA COLLECTION        : 12-MAY-1995
REMARK 200  TEMPERATURE           (KELVIN) : 120
REMARK 200  PH                             : 7.5
REMARK 200  NUMBER OF CRYSTALS USED        : 1
REMARK 200
REMARK 200  SYNCHROTRON              (Y/N) : Y
REMARK 200  RADIATION SOURCE               : EMBL/DESY, HAMBURG
REMARK 200  BEAMLINE                       : X31
REMARK 200  X-RAY GENERATOR MODEL          : NULL
REMARK 200  MONOCHROMATIC OR LAUE    (M/L) : M
REMARK 200  WAVELENGTH OR RANGE        (A) : 0.92
REMARK 200  MONOCHROMATOR                  : NULL
REMARK 200  OPTICS                         : NULL
REMARK 200
REMARK 200  DETECTOR TYPE                  : IMAGE PLATE
REMARK 200  DETECTOR MANUFACTURER          : MARRESEARCH
REMARK 200  INTENSITY-INTEGRATION SOFTWARE : DENZO
REMARK 200  DATA SCALING SOFTWARE          : SCALEPACK
REMARK 200
REMARK 200  NUMBER OF UNIQUE REFLECTIONS   : 24031
REMARK 200  RESOLUTION RANGE HIGH      (A) : 1.90
REMARK 200  RESOLUTION RANGE LOW       (A) : 15.0
REMARK 200  REJECTION CRITERIA  (SIGMA(I)) : 0.0
REMARK 200
REMARK 200 OVERALL.
REMARK 200  COMPLETENESS FOR RANGE     (%) : 90.4
REMARK 200  DATA REDUNDANCY                : 5.0
REMARK 200  R MERGE                    (I) : 0.073
REMARK 200  R SYM                      (I) : NULL
REMARK 200  <I/SIGMA(I)> FOR THE DATA SET  : 0.0
REMARK 200
REMARK 200 IN THE HIGHEST RESOLUTION SHELL.
REMARK 200  HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.90
REMARK 200  HIGHEST RESOLUTION SHELL, RANGE LOW  (A) : 1.93
REMARK 200  COMPLETENESS FOR SHELL     (%) : 67.8
REMARK 200  DATA REDUNDANCY IN SHELL       : 2.5
REMARK 200  R MERGE FOR SHELL          (I) : 0.355
REMARK 200  R SYM FOR SHELL            (I) : NULL
REMARK 200  <I/SIGMA(I)> FOR SHELL         : 0.0
REMARK 200
REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MIR
REMARK 200 SOFTWARE USED: CCP4
REMARK 200 STARTING MODEL: NULL
REMARK 200
REMARK 200 REMARK: NULL
REMARK 280
REMARK 280 CRYSTAL
REMARK 280 SOLVENT CONTENT, VS   (%): 56.
REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.8
REMARK 280
REMARK 280 CRYSTALLIZATION CONDITIONS: 0.1 M NATRIUM HEPES BUFFER
REMARK 280 PH=7.5 2 % PEG 400, 2.0 M AMMONIUM SULFATE, 20 DEGREES.
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY
REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: C 2 2 21
REMARK 290
REMARK 290      SYMOP   SYMMETRY
REMARK 290     NNNMMM   OPERATOR
REMARK 290       1555   X,Y,Z
REMARK 290       2555   -X,-Y,1/2+Z
REMARK 290       3555   -X,Y,1/2-Z
REMARK 290       4555   X,-Y,-Z
REMARK 290       5555   1/2+X,1/2+Y,Z
REMARK 290       6555   1/2-X,1/2-Y,1/2+Z
REMARK 290       7555   1/2-X,1/2+Y,1/2-Z
REMARK 290       8555   1/2+X,1/2-Y,-Z
REMARK 290
REMARK 290     WHERE NNN -> OPERATOR NUMBER
REMARK 290           MMM -> TRANSLATION VECTOR
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS
REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM
REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY
REMARK 290 RELATED MOLECULES.
REMARK 290   SMTRY1   1  1.000000  0.000000  0.000000        0.00000
REMARK 290   SMTRY2   1  0.000000  1.000000  0.000000        0.00000
REMARK 290   SMTRY3   1  0.000000  0.000000  1.000000        0.00000
REMARK 290   SMTRY1   2 -1.000000  0.000000  0.000000        0.00000
REMARK 290   SMTRY2   2  0.000000 -1.000000  0.000000        0.00000
REMARK 290   SMTRY3   2  0.000000  0.000000  1.000000       64.11079
REMARK 290   SMTRY1   3 -1.000000  0.000000  0.000000        0.00000
REMARK 290   SMTRY2   3  0.000000  1.000000  0.000000        0.00000
REMARK 290   SMTRY3   3  0.000000  0.000000 -1.000000       64.11079
REMARK 290   SMTRY1   4  1.000000  0.000000  0.000000        0.00000
REMARK 290   SMTRY2   4  0.000000 -1.000000  0.000000        0.00000
REMARK 290   SMTRY3   4  0.000000  0.000000 -1.000000        0.00000
REMARK 290   SMTRY1   5  1.000000  0.000000  0.000000       23.85155
REMARK 290   SMTRY2   5  0.000000  1.000000  0.000000       54.50779
REMARK 290   SMTRY3   5  0.000000  0.000000  1.000000        0.00000
REMARK 290   SMTRY1   6 -1.000000  0.000000  0.000000       23.85155
REMARK 290   SMTRY2   6  0.000000 -1.000000  0.000000       54.50779
REMARK 290   SMTRY3   6  0.000000  0.000000  1.000000       64.11079
REMARK 290   SMTRY1   7 -1.000000  0.000000  0.000000       23.85155
REMARK 290   SMTRY2   7  0.000000  1.000000  0.000000       54.50779
REMARK 290   SMTRY3   7  0.000000  0.000000 -1.000000       64.11079
REMARK 290   SMTRY1   8  1.000000  0.000000  0.000000       23.85155
REMARK 290   SMTRY2   8  0.000000 -1.000000  0.000000       54.50779
REMARK 290   SMTRY3   8  0.000000  0.000000 -1.000000        0.00000
REMARK 290
REMARK 290 REMARK: NULL
REMARK 375
REMARK 375 SPECIAL POSITION
REMARK 375      HOH   575  LIES ON A SPECIAL POSITION.
REMARK 375      HOH   576  LIES ON A SPECIAL POSITION.
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: CLOSE CONTACTS
REMARK 500
REMARK 500 THE FOLLOWING ATOMS THAT ARE RELATED BY CRYSTALLOGRAPHIC
REMARK 500 SYMMETRY ARE IN CLOSE CONTACT.  SOME OF THESE MAY BE ATOMS
REMARK 500 LOCATED ON SPECIAL POSITIONS IN THE CELL.  ATOMS WITH
REMARK 500 NON-BLANK ALTERNATE LOCATION INDICATORS ARE NOT INCLUDED
REMARK 500 IN THE CALCULATIONS.
REMARK 500
REMARK 500 DISTANCE CUTOFF:
REMARK 500 2.2 ANGSTROMS FOR CONTACTS NOT INVOLVING HYDROGEN ATOMS
REMARK 500 1.6 ANGSTROMS FOR CONTACTS INVOLVING HYDROGEN ATOMS
REMARK 500
REMARK 500  ATM1  RES C  SSEQI   ATM2  RES C  SSEQI  SSYMOP   DISTANCE
REMARK 500   O    HOH     575     O    HOH     575     3655     0.50
REMARK 500   O    HOH     576     O    HOH     576     3655     0.78
REMARK 800
REMARK 800 SITE
REMARK 800 SITE_IDENTIFIER: CAT
REMARK 800 SITE_DESCRIPTION: BINDING SITE.
REMARK 999
REMARK 999 SEQUENCE
REMARK 999 1YAS  A    SWS     P52704       1 -     1 NOT IN ATOMS LIST
DBREF  1YAS A    2   257  SWS    P52704   HNL_HEVBR        2    257
DBREF  1YAS B  300   300  PDB    1YAS     1YAS           300    300
SEQRES   1 A  257  MET ALA PHE ALA HIS PHE VAL LEU ILE HIS THR ILE CYS
SEQRES   2 A  257  HIS GLY ALA TRP ILE TRP HIS LYS LEU LYS PRO LEU LEU
SEQRES   3 A  257  GLU ALA LEU GLY HIS LYS VAL THR ALA LEU ASP LEU ALA
SEQRES   4 A  257  ALA SER GLY VAL ASP PRO ARG GLN ILE GLU GLU ILE GLY
SEQRES   5 A  257  SER PHE ASP GLU TYR SER GLU PRO LEU LEU THR PHE LEU
SEQRES   6 A  257  GLU ALA LEU PRO PRO GLY GLU LYS VAL ILE LEU VAL GLY
SEQRES   7 A  257  GLU SER CYS GLY GLY LEU ASN ILE ALA ILE ALA ALA ASP
SEQRES   8 A  257  LYS TYR CYS GLU LYS ILE ALA ALA ALA VAL PHE HIS ASN
SEQRES   9 A  257  SER VAL LEU PRO ASP THR GLU HIS CYS PRO SER TYR VAL
SEQRES  10 A  257  VAL ASP LYS LEU MET GLU VAL PHE PRO ASP TRP LYS ASP
SEQRES  11 A  257  THR THR TYR PHE THR TYR THR LYS ASP GLY LYS GLU ILE
SEQRES  12 A  257  THR GLY LEU LYS LEU GLY PHE THR LEU LEU ARG GLU ASN
SEQRES  13 A  257  LEU TYR THR LEU CYS GLY PRO GLU GLU TYR GLU LEU ALA
SEQRES  14 A  257  LYS MET LEU THR ARG LYS GLY SER LEU PHE GLN ASN ILE
SEQRES  15 A  257  LEU ALA LYS ARG PRO PHE PHE THR LYS GLU GLY TYR GLY
SEQRES  16 A  257  SER ILE LYS LYS ILE TYR VAL TRP THR ASP GLN ASP GLU
SEQRES  17 A  257  ILE PHE LEU PRO GLU PHE GLN LEU TRP GLN ILE GLU ASN
SEQRES  18 A  257  TYR LYS PRO ASP LYS VAL TYR LYS VAL GLU GLY GLY ASP
SEQRES  19 A  257  HIS LYS LEU GLN LEU THR LYS THR LYS GLU ILE ALA GLU
SEQRES  20 A  257  ILE LEU GLN GLU VAL ALA ASP THR TYR ASN
SEQRES   1 B    1  HIS
HET    SO4    301       5
HETNAM     SO4 SULFATE ION
FORMUL   3  SO4    O4 S1 2-
FORMUL   4  HOH   *77(H2 O1)
HELIX    1  HA LYS A   21  ALA A   28  1                                   8
HELIX    2  HB SER A   53  LEU A   68  1                                  16
HELIX    3  HC CYS A   81  CYS A   94  1                                  14
HELIX    4 HD1 SER A  115  PHE A  125  1 BELONGS TO THE CAP DOMAIN        11
HELIX    5 HD2 GLY A  149  LEU A  157  1 BELONGS TO THE CAP DOMAIN         9
HELIX    6 HD3 GLY A  162  THR A  173  1 BELONGS TO THE CAP DOMAIN        12
HELIX    7  HD PHE A  179  LYS A  185  1                                   7
HELIX    8  HE LEU A  211  TYR A  222  1                                  12
HELIX    9  HF LYS A  236  TYR A  256  1                                  21
SHEET    1  S1 6 ALA A   4  ILE A   9  0
SHEET    2  S1 6 LYS A  32  LEU A  36  1
SHEET    3  S1 6 SER A  53  LEU A  68 -1
SHEET    4  S1 6 LYS A  96  ASN A 104 -1
SHEET    5  S1 6 LYS A 198  TRP A 203  1
SHEET    6  S1 6 LYS A 226  VAL A 230  1
SHEET    1  S2 3 THR A 132  LYS A 138  0
SHEET    2  S2 3 LYS A 141  LYS A 147 -1
SHEET    3  S2 3 GLY A 176  LEU A 178 -1
SITE     1 CAT  1 HIS A 235
CRYST1   47.703  109.010  128.226  90.00  90.00  90.00 C 2 2 21      8
ORIGX1      1.000000  0.000000  0.000000        0.00000
ORIGX2      0.000000  1.000000  0.000000        0.00000
ORIGX3      0.000000  0.000000  1.000000        0.00000
SCALE1      0.020963  0.000000  0.000000        0.00000
SCALE2      0.000000  0.009173  0.000000        0.00000
SCALE3      0.000000  0.000000  0.007799        0.00000