longtext: 4ufn-pdb

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HEADER    HYDROLASE                               17-MAR-15   4UFN
TITLE     LABORATORY EVOLVED VARIANT R-C1B1 OF POTATO EPOXIDE
TITLE    2 HYDROLASE STEH1
COMPND    MOL_ID: 1;
COMPND   2 MOLECULE: EPOXIDE HYDROLASE;
COMPND   3 CHAIN: A, B;
COMPND   4 FRAGMENT: RESIDUES 3-321;
COMPND   5 EC: 3.3.2.3;
COMPND   6 ENGINEERED: YES;
COMPND   7 MUTATION: YES
SOURCE    MOL_ID: 1;
SOURCE   2 ORGANISM_SCIENTIFIC: SOLANUM TUBEROSUM;
SOURCE   3 ORGANISM_COMMON: POTATO;
SOURCE   4 ORGANISM_TAXID: 4113;
SOURCE   5 STRAIN: LEMHI RUSSET;
SOURCE   6 EXPRESSION_SYSTEM: ESCHERICHIA COLI;
SOURCE   7 EXPRESSION_SYSTEM_TAXID: 562;
SOURCE   8 EXPRESSION_SYSTEM_STRAIN: XL1 BLUE;
SOURCE   9 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID;
SOURCE  10 EXPRESSION_SYSTEM_PLASMID: PGTACSTEH1-5H
KEYWDS    HYDROLASE, EPOXIDE HYDROLYSIS, ALPHA/BETA-HYDROLASE, DIRECTED
KEYWDS   2 EVOLUTION, ASYMMETRIC SYNTHESES
EXPDTA    X-RAY DIFFRACTION
AUTHOR    A.J.CARLSSON,P.BAUER,M.NILSSON,D.DOBRITZSCH,S.C.L.KAMERLIN,
AUTHOR   2 M.WIDERSTEN
REVDAT   1   13-APR-16 4UFN    0
JRNL        AUTH   A.J.CARLSSON,P.BAUER,M.NILSSON,D.DOBRITZSCH,S.C.L.KAMERLIN,
JRNL        AUTH 2 M.WIDERSTEN
JRNL        TITL   PROBING INFLUENCE OF ACTIVE-SITE VOLUME TO CATALYSIS:
JRNL        TITL 2 LABORATORY EVOLVED EPOXIDE HYDROLASES AS MODEL ENZYMES
JRNL        REF    TO BE PUBLISHED
JRNL        REFN
REMARK   2
REMARK   2 RESOLUTION.    2.00 ANGSTROMS.
REMARK   3
REMARK   3 REFINEMENT.
REMARK   3   PROGRAM     : REFMAC 5.8.0069
REMARK   3   AUTHORS     : MURSHUDOV,SKUBAK,LEBEDEV,PANNU,
REMARK   3                 STEINER,NICHOLLS,WINN,LONG,VAGIN
REMARK   3
REMARK   3    REFINEMENT TARGET : MAXIMUM LIKELIHOOD
REMARK   3
REMARK   3  DATA USED IN REFINEMENT.
REMARK   3   RESOLUTION RANGE HIGH (ANGSTROMS) : 2.00
REMARK   3   RESOLUTION RANGE LOW  (ANGSTROMS) : 77.30
REMARK   3   DATA CUTOFF            (SIGMA(F)) : NULL
REMARK   3   COMPLETENESS FOR RANGE        (%) : 99.22
REMARK   3   NUMBER OF REFLECTIONS             : 44563
REMARK   3
REMARK   3  FIT TO DATA USED IN REFINEMENT.
REMARK   3   CROSS-VALIDATION METHOD          : THROUGHOUT
REMARK   3   FREE R VALUE TEST SET SELECTION  : RANDOM
REMARK   3   R VALUE     (WORKING + TEST SET) : 0.17255
REMARK   3   R VALUE            (WORKING SET) : 0.17055
REMARK   3   FREE R VALUE                     : 0.21111
REMARK   3   FREE R VALUE TEST SET SIZE   (%) : 4.9
REMARK   3   FREE R VALUE TEST SET COUNT      : 2292
REMARK   3
REMARK   3  FIT IN THE HIGHEST RESOLUTION BIN.
REMARK   3   TOTAL NUMBER OF BINS USED           : 20
REMARK   3   BIN RESOLUTION RANGE HIGH       (A) : 2.000
REMARK   3   BIN RESOLUTION RANGE LOW        (A) : 2.052
REMARK   3   REFLECTION IN BIN     (WORKING SET) : 3231
REMARK   3   BIN COMPLETENESS (WORKING+TEST) (%) : 99.18
REMARK   3   BIN R VALUE           (WORKING SET) : 0.232
REMARK   3   BIN FREE R VALUE SET COUNT          : 147
REMARK   3   BIN FREE R VALUE                    : 0.267
REMARK   3
REMARK   3  NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
REMARK   3   PROTEIN ATOMS            : 5126
REMARK   3   NUCLEIC ACID ATOMS       : 0
REMARK   3   HETEROGEN ATOMS          : 12
REMARK   3   SOLVENT ATOMS            : 642
REMARK   3
REMARK   3  B VALUES.
REMARK   3   FROM WILSON PLOT           (A**2) : 20.0
REMARK   3   MEAN B VALUE      (OVERALL, A**2) : 26.371
REMARK   3   OVERALL ANISOTROPIC B VALUE.
REMARK   3    B11 (A**2) : 1.58
REMARK   3    B22 (A**2) : 0.08
REMARK   3    B33 (A**2) : -1.66
REMARK   3    B12 (A**2) : 0.00
REMARK   3    B13 (A**2) : 0.00
REMARK   3    B23 (A**2) : 0.00
REMARK   3
REMARK   3  ESTIMATED OVERALL COORDINATE ERROR.
REMARK   3   ESU BASED ON R VALUE                            (A): 0.182
REMARK   3   ESU BASED ON FREE R VALUE                       (A): 0.155
REMARK   3   ESU BASED ON MAXIMUM LIKELIHOOD                 (A): 0.112
REMARK   3   ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 4.132
REMARK   3
REMARK   3 CORRELATION COEFFICIENTS.
REMARK   3   CORRELATION COEFFICIENT FO-FC      : 0.959
REMARK   3   CORRELATION COEFFICIENT FO-FC FREE : 0.937
REMARK   3
REMARK   3  RMS DEVIATIONS FROM IDEAL VALUES        COUNT    RMS    WEIGHT
REMARK   3   BOND LENGTHS REFINED ATOMS        (A):  5334 ; 0.009 ; 0.019
REMARK   3   BOND LENGTHS OTHERS               (A):  5090 ; 0.004 ; 0.020
REMARK   3   BOND ANGLES REFINED ATOMS   (DEGREES):  7253 ; 1.304 ; 1.956
REMARK   3   BOND ANGLES OTHERS          (DEGREES): 11747 ; 1.002 ; 3.000
REMARK   3   TORSION ANGLES, PERIOD 1    (DEGREES):   650 ; 5.821 ; 5.000
REMARK   3   TORSION ANGLES, PERIOD 2    (DEGREES):   243 ;31.581 ;23.909
REMARK   3   TORSION ANGLES, PERIOD 3    (DEGREES):   870 ;13.304 ;15.000
REMARK   3   TORSION ANGLES, PERIOD 4    (DEGREES):    20 ;17.478 ;15.000
REMARK   3   CHIRAL-CENTER RESTRAINTS       (A**3):   770 ; 0.076 ; 0.200
REMARK   3   GENERAL PLANES REFINED ATOMS      (A):  5980 ; 0.007 ; 0.021
REMARK   3   GENERAL PLANES OTHERS             (A):  1244 ; 0.004 ; 0.020
REMARK   3   NON-BONDED CONTACTS REFINED ATOMS (A):  NULL ;  NULL ;  NULL
REMARK   3   NON-BONDED CONTACTS OTHERS        (A):  NULL ;  NULL ;  NULL
REMARK   3   NON-BONDED TORSION REFINED ATOMS  (A):  NULL ;  NULL ;  NULL
REMARK   3   NON-BONDED TORSION OTHERS         (A):  NULL ;  NULL ;  NULL
REMARK   3   H-BOND (X...Y) REFINED ATOMS      (A):  NULL ;  NULL ;  NULL
REMARK   3   H-BOND (X...Y) OTHERS             (A):  NULL ;  NULL ;  NULL
REMARK   3   POTENTIAL METAL-ION REFINED ATOMS (A):  NULL ;  NULL ;  NULL
REMARK   3   POTENTIAL METAL-ION OTHERS        (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY VDW REFINED ATOMS        (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY VDW OTHERS               (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY H-BOND REFINED ATOMS     (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY H-BOND OTHERS            (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY METAL-ION REFINED ATOMS  (A):  NULL ;  NULL ;  NULL
REMARK   3   SYMMETRY METAL-ION OTHERS         (A):  NULL ;  NULL ;  NULL
REMARK   3
REMARK   3  ISOTROPIC THERMAL FACTOR RESTRAINTS.     COUNT   RMS    WEIGHT
REMARK   3   MAIN-CHAIN BOND REFINED ATOMS  (A**2):  2570 ; 1.516 ; 2.436
REMARK   3   MAIN-CHAIN BOND OTHER ATOMS    (A**2):  2569 ; 1.515 ; 2.436
REMARK   3   MAIN-CHAIN ANGLE REFINED ATOMS (A**2):  3212 ; 2.393 ; 3.644
REMARK   3   MAIN-CHAIN ANGLE OTHER ATOMS   (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SIDE-CHAIN BOND REFINED ATOMS  (A**2):  2764 ; 1.948 ; 2.632
REMARK   3   SIDE-CHAIN BOND OTHER ATOMS    (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SIDE-CHAIN ANGLE REFINED ATOMS (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SIDE-CHAIN ANGLE OTHER ATOMS   (A**2):  NULL ;  NULL ;  NULL
REMARK   3   LONG RANGE B REFINED ATOMS     (A**2):  NULL ;  NULL ;  NULL
REMARK   3   LONG RANGE B OTHER ATOMS       (A**2):  NULL ;  NULL ;  NULL
REMARK   3
REMARK   3 ANISOTROPIC THERMAL FACTOR RESTRAINTS.    COUNT   RMS   WEIGHT
REMARK   3   RIGID-BOND RESTRAINTS          (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SPHERICITY; FREE ATOMS         (A**2):  NULL ;  NULL ;  NULL
REMARK   3   SPHERICITY; BONDED ATOMS       (A**2):  NULL ;  NULL ;  NULL
REMARK   3
REMARK   3  NCS RESTRAINTS STATISTICS
REMARK   3   NCS TYPE: LOCAL
REMARK   3   NUMBER OF DIFFERENT NCS PAIRS  : 1
REMARK   3  GROUP  CHAIN1    RANGE     CHAIN2     RANGE    COUNT RMS  WEIGHT
REMARK   3    1      A     2   321       B     2    321    19314  0.11  0.05
REMARK   3
REMARK   3  TLS DETAILS
REMARK   3   NUMBER OF TLS GROUPS  : NULL
REMARK   3
REMARK   3  BULK SOLVENT MODELLING.
REMARK   3   METHOD USED : MASK
REMARK   3   PARAMETERS FOR MASK CALCULATION
REMARK   3   VDW PROBE RADIUS   : 1.20
REMARK   3   ION PROBE RADIUS   : 0.80
REMARK   3   SHRINKAGE RADIUS   : 0.80
REMARK   3
REMARK   3  OTHER REFINEMENT REMARKS: HYDROGENS HAVE BEEN ADDED IN THE RIDING
REMARK   3  POSITIONS. U VALUES REFINED INDIVIDUALLY
REMARK   4
REMARK   4 4UFN COMPLIES WITH FORMAT V. 3.30, 13-JUL-11
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE ON 17-MAR-15.
REMARK 100 THE PDBE ID CODE IS EBI-63367.
REMARK 200
REMARK 200 EXPERIMENTAL DETAILS
REMARK 200  EXPERIMENT TYPE                : X-RAY DIFFRACTION
REMARK 200  DATE OF DATA COLLECTION        : 10-MAY-14
REMARK 200  TEMPERATURE           (KELVIN) : 100
REMARK 200  PH                             : 8.0
REMARK 200  NUMBER OF CRYSTALS USED        : 1
REMARK 200
REMARK 200  SYNCHROTRON              (Y/N) : Y
REMARK 200  RADIATION SOURCE               : DIAMOND
REMARK 200  BEAMLINE                       : I04
REMARK 200  X-RAY GENERATOR MODEL          : NULL
REMARK 200  MONOCHROMATIC OR LAUE    (M/L) : M
REMARK 200  WAVELENGTH OR RANGE        (A) : 0.9763
REMARK 200  MONOCHROMATOR                  : NULL
REMARK 200  OPTICS                         : NULL
REMARK 200
REMARK 200  DETECTOR TYPE                  : PIXEL (PILATUS 6M)
REMARK 200  DETECTOR MANUFACTURER          : DECTRIS
REMARK 200  INTENSITY-INTEGRATION SOFTWARE : XDS
REMARK 200  DATA SCALING SOFTWARE          : AIMLESS
REMARK 200
REMARK 200  NUMBER OF UNIQUE REFLECTIONS   : 47143
REMARK 200  RESOLUTION RANGE HIGH      (A) : 2.00
REMARK 200  RESOLUTION RANGE LOW       (A) : 99.14
REMARK 200  REJECTION CRITERIA  (SIGMA(I)) : 2.0
REMARK 200
REMARK 200 OVERALL.
REMARK 200  COMPLETENESS FOR RANGE     (%) : 99.9
REMARK 200  DATA REDUNDANCY                : 5.7
REMARK 200  R MERGE                    (I) : 0.12
REMARK 200  R SYM                      (I) : NULL
REMARK 200   FOR THE DATA SET  : 9.00
REMARK 200
REMARK 200 IN THE HIGHEST RESOLUTION SHELL.
REMARK 200  HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.00
REMARK 200  HIGHEST RESOLUTION SHELL, RANGE LOW  (A) : 2.05
REMARK 200  COMPLETENESS FOR SHELL     (%) : 100.0
REMARK 200  DATA REDUNDANCY IN SHELL       : 5.8
REMARK 200  R MERGE FOR SHELL          (I) : 0.57
REMARK 200  R SYM FOR SHELL            (I) : NULL
REMARK 200   FOR SHELL         : 2.60
REMARK 200
REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH
REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT
REMARK 200 SOFTWARE USED: PHASER
REMARK 200 STARTING MODEL: PDB ENTRY 2CJP
REMARK 200
REMARK 200 REMARK: NONE
REMARK 280
REMARK 280 CRYSTAL
REMARK 280 SOLVENT CONTENT, VS   (%): 46
REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.3
REMARK 280
REMARK 280 CRYSTALLIZATION CONDITIONS: 18% (W/V) PEG5K-MME 0.1 M TRIS PH 8.0
REMARK 280  5% (V/V) 1,4-DIOXANE
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY
REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 21
REMARK 290
REMARK 290      SYMOP   SYMMETRY
REMARK 290     NNNMMM   OPERATOR
REMARK 290       1555   X,Y,Z
REMARK 290       2555   -X+1/2,-Y,Z+1/2
REMARK 290       3555   -X,Y+1/2,-Z+1/2
REMARK 290       4555   X+1/2,-Y+1/2,-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       28.00150
REMARK 290   SMTRY2   2  0.000000 -1.000000  0.000000        0.00000
REMARK 290   SMTRY3   2  0.000000  0.000000  1.000000       61.73050
REMARK 290   SMTRY1   3 -1.000000  0.000000  0.000000        0.00000
REMARK 290   SMTRY2   3  0.000000  1.000000  0.000000       49.57000
REMARK 290   SMTRY3   3  0.000000  0.000000 -1.000000       61.73050
REMARK 290   SMTRY1   4  1.000000  0.000000  0.000000       28.00150
REMARK 290   SMTRY2   4  0.000000 -1.000000  0.000000       49.57000
REMARK 290   SMTRY3   4  0.000000  0.000000 -1.000000        0.00000
REMARK 290
REMARK 290 REMARK: NULL
REMARK 300
REMARK 300 BIOMOLECULE: 1, 2
REMARK 300 SEE REMARK 350 FOR THE AUTHOR PROVIDED AND/OR PROGRAM
REMARK 300 GENERATED ASSEMBLY INFORMATION FOR THE STRUCTURE IN
REMARK 300 THIS ENTRY. THE REMARK MAY ALSO PROVIDE INFORMATION ON
REMARK 300 BURIED SURFACE AREA.
REMARK 350
REMARK 350 COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN
REMARK 350 BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE
REMARK 350 MOLECULE CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS
REMARK 350 GIVEN BELOW.  BOTH NON-CRYSTALLOGRAPHIC AND
REMARK 350 CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN.
REMARK 350
REMARK 350 BIOMOLECULE: 1
REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: MONOMERIC
REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: MONOMERIC
REMARK 350 SOFTWARE USED: PQS
REMARK 350 APPLY THE FOLLOWING TO CHAINS: A
REMARK 350   BIOMT1   1  1.000000  0.000000  0.000000        0.00000
REMARK 350   BIOMT2   1  0.000000  1.000000  0.000000        0.00000
REMARK 350   BIOMT3   1  0.000000  0.000000  1.000000        0.00000
REMARK 350
REMARK 350 BIOMOLECULE: 2
REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: MONOMERIC
REMARK 350 SOFTWARE DETERMINED QUATERNARY STRUCTURE: MONOMERIC
REMARK 350 SOFTWARE USED: PQS
REMARK 350 APPLY THE FOLLOWING TO CHAINS: B
REMARK 350   BIOMT1   1  1.000000  0.000000  0.000000        0.00000
REMARK 350   BIOMT2   1  0.000000  1.000000  0.000000        0.00000
REMARK 350   BIOMT3   1  0.000000  0.000000  1.000000        0.00000
REMARK 465
REMARK 465 MISSING RESIDUES
REMARK 465 THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE
REMARK 465 EXPERIMENT. (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
REMARK 465 IDENTIFIER; SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.)
REMARK 465
REMARK 465   M RES C SSSEQI
REMARK 465     MET A     1
REMARK 465     SER A   323
REMARK 465     HIS A   324
REMARK 465     HIS A   325
REMARK 465     HIS A   326
REMARK 465     HIS A   327
REMARK 465     HIS A   328
REMARK 465     MET B     1
REMARK 465     SER B   323
REMARK 465     HIS B   324
REMARK 465     HIS B   325
REMARK 465     HIS B   326
REMARK 465     HIS B   327
REMARK 465     HIS B   328
REMARK 470
REMARK 470 MISSING ATOM
REMARK 470 THE FOLLOWING RESIDUES HAVE MISSING ATOMS (M=MODEL NUMBER;
REMARK 470 RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE NUMBER;
REMARK 470 I=INSERTION CODE):
REMARK 470   M RES CSSEQI  ATOMS
REMARK 470     THR A 322    CA   C    O    CB   OG1  CG2
REMARK 470     THR B 322    CA   C    O    CB   OG1  CG2
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: CLOSE CONTACTS IN SAME ASYMMETRIC UNIT
REMARK 500
REMARK 500 THE FOLLOWING ATOMS ARE IN CLOSE CONTACT.
REMARK 500
REMARK 500  ATM1  RES C  SSEQI   ATM2  RES C  SSEQI           DISTANCE
REMARK 500   OE1  GLU A    90     O    HOH A  2119              2.11
REMARK 500
REMARK 500 REMARK: NULL
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: TORSION ANGLES
REMARK 500
REMARK 500 TORSION ANGLES OUTSIDE THE EXPECTED RAMACHANDRAN REGIONS:
REMARK 500 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER;
REMARK 500 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE).
REMARK 500
REMARK 500 STANDARD TABLE:
REMARK 500 FORMAT:(10X,I3,1X,A3,1X,A1,I4,A1,4X,F7.2,3X,F7.2)
REMARK 500
REMARK 500 EXPECTED VALUES: GJ KLEYWEGT AND TA JONES (1996). PHI/PSI-
REMARK 500 CHOLOGY: RAMACHANDRAN REVISITED. STRUCTURE 4, 1395 - 1400
REMARK 500
REMARK 500  M RES CSSEQI        PSI       PHI
REMARK 500    GLU A  35     -161.57   -121.33
REMARK 500    ASP A 105     -123.98     64.94
REMARK 500    SER A 129      -52.48     80.10
REMARK 500    ALA A 299     -141.01   -110.84
REMARK 500    GLU B  35     -161.44   -121.60
REMARK 500    ASP B 105     -124.57     63.56
REMARK 500    SER B 129      -58.34     78.73
REMARK 500    ALA B 299     -143.61   -112.04
REMARK 500    PHE B 301       59.99    -92.40
REMARK 500
REMARK 500 REMARK: NULL
REMARK 800
REMARK 800 SITE
REMARK 800 SITE_IDENTIFIER: AC1
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE DIO A1322
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC2
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE DIO B1322
REMARK 900
REMARK 900 RELATED ENTRIES
REMARK 900 RELATED ID: 4UFO   RELATED DB: PDB
REMARK 900  LABORATORY EVOLVED VARIANT R-C1B1D33E6 OF POTATO EPOXIDE
REMARK 900   HYDROLASE STEH1
REMARK 900 RELATED ID: 4UFP   RELATED DB: PDB
REMARK 900  LABORATORY EVOLVED VARIANT R-C1B1D33 OF POTATO EPOXIDE
REMARK 900  HYDROLASE STEH1
REMARK 999
REMARK 999 SEQUENCE
REMARK 999 THE SEQUENCE IS N-TERMINALLY HIS-TAGGED AND IS MUTATED AT
REMARK 999 5 POSITIONS (P84L, W106L, L109Y, V141K, I155V)
DBREF  4UFN A    1   321  UNP    Q41415   Q41415_SOLTU     1    321
DBREF  4UFN B    1   321  UNP    Q41415   Q41415_SOLTU     1    321
SEQADV 4UFN LYS A    2  UNP  Q41415    GLU     2 CLONING ARTIFACT
SEQADV 4UFN LEU A   94  UNP  Q41415    PRO    94 ENGINEERED MUTATION
SEQADV 4UFN LEU A  106  UNP  Q41415    TRP   106 ENGINEERED MUTATION
SEQADV 4UFN TYR A  109  UNP  Q41415    LEU   109 ENGINEERED MUTATION
SEQADV 4UFN LYS A  141  UNP  Q41415    VAL   141 ENGINEERED MUTATION
SEQADV 4UFN VAL A  155  UNP  Q41415    ILE   155 ENGINEERED MUTATION
SEQADV 4UFN THR A  322  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN SER A  323  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS A  324  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS A  325  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS A  326  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS A  327  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS A  328  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN LYS B    2  UNP  Q41415    GLU     2 CLONING ARTIFACT
SEQADV 4UFN LEU B   94  UNP  Q41415    PRO    94 ENGINEERED MUTATION
SEQADV 4UFN LEU B  106  UNP  Q41415    TRP   106 ENGINEERED MUTATION
SEQADV 4UFN TYR B  109  UNP  Q41415    LEU   109 ENGINEERED MUTATION
SEQADV 4UFN LYS B  141  UNP  Q41415    VAL   141 ENGINEERED MUTATION
SEQADV 4UFN VAL B  155  UNP  Q41415    ILE   155 ENGINEERED MUTATION
SEQADV 4UFN THR B  322  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN SER B  323  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS B  324  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS B  325  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS B  326  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS B  327  UNP  Q41415              EXPRESSION TAG
SEQADV 4UFN HIS B  328  UNP  Q41415              EXPRESSION TAG
SEQRES   1 A  328  MET LYS LYS ILE GLU HIS LYS MET VAL ALA VAL ASN GLY
SEQRES   2 A  328  LEU ASN MET HIS LEU ALA GLU LEU GLY GLU GLY PRO THR
SEQRES   3 A  328  ILE LEU PHE ILE HIS GLY PHE PRO GLU LEU TRP TYR SER
SEQRES   4 A  328  TRP ARG HIS GLN MET VAL TYR LEU ALA GLU ARG GLY TYR
SEQRES   5 A  328  ARG ALA VAL ALA PRO ASP LEU ARG GLY TYR GLY ASP THR
SEQRES   6 A  328  THR GLY ALA PRO LEU ASN ASP PRO SER LYS PHE SER ILE
SEQRES   7 A  328  LEU HIS LEU VAL GLY ASP VAL VAL ALA LEU LEU GLU ALA
SEQRES   8 A  328  ILE ALA LEU ASN GLU GLU LYS VAL PHE VAL VAL ALA HIS
SEQRES   9 A  328  ASP LEU GLY ALA TYR ILE ALA TRP HIS LEU CYS LEU PHE
SEQRES  10 A  328  ARG PRO ASP LYS VAL LYS ALA LEU VAL ASN LEU SER VAL
SEQRES  11 A  328  HIS PHE SER LYS ARG ASN PRO LYS MET ASN LYS VAL GLU
SEQRES  12 A  328  GLY LEU LYS ALA ILE TYR GLY GLU ASP HIS TYR VAL SER
SEQRES  13 A  328  ARG PHE GLN VAL PRO GLY GLU ILE GLU ALA GLU PHE ALA
SEQRES  14 A  328  PRO ILE GLY ALA LYS SER VAL LEU LYS LYS ILE LEU THR
SEQRES  15 A  328  TYR ARG ASP PRO ALA PRO PHE TYR PHE PRO LYS GLY LYS
SEQRES  16 A  328  GLY LEU GLU ALA ILE PRO ASP ALA PRO VAL ALA LEU SER
SEQRES  17 A  328  SER TRP LEU SER GLU GLU GLU LEU ASP TYR TYR ALA ASN
SEQRES  18 A  328  LYS PHE GLU GLN THR GLY PHE THR GLY ALA VAL ASN TYR
SEQRES  19 A  328  TYR ARG ALA LEU PRO ILE ASN TRP GLU LEU THR ALA PRO
SEQRES  20 A  328  TRP THR GLY ALA GLN VAL LYS VAL PRO THR LYS PHE ILE
SEQRES  21 A  328  VAL GLY GLU PHE ASP LEU VAL TYR HIS ILE PRO GLY ALA
SEQRES  22 A  328  LYS GLU TYR ILE HIS ASN GLY GLY PHE LYS LYS ASP VAL
SEQRES  23 A  328  PRO LEU LEU GLU GLU VAL VAL VAL LEU GLU GLY ALA ALA
SEQRES  24 A  328  HIS PHE VAL SER GLN GLU ARG PRO HIS GLU ILE SER LYS
SEQRES  25 A  328  HIS ILE TYR ASP PHE ILE GLN LYS PHE THR SER HIS HIS
SEQRES  26 A  328  HIS HIS HIS
SEQRES   1 B  328  MET LYS LYS ILE GLU HIS LYS MET VAL ALA VAL ASN GLY
SEQRES   2 B  328  LEU ASN MET HIS LEU ALA GLU LEU GLY GLU GLY PRO THR
SEQRES   3 B  328  ILE LEU PHE ILE HIS GLY PHE PRO GLU LEU TRP TYR SER
SEQRES   4 B  328  TRP ARG HIS GLN MET VAL TYR LEU ALA GLU ARG GLY TYR
SEQRES   5 B  328  ARG ALA VAL ALA PRO ASP LEU ARG GLY TYR GLY ASP THR
SEQRES   6 B  328  THR GLY ALA PRO LEU ASN ASP PRO SER LYS PHE SER ILE
SEQRES   7 B  328  LEU HIS LEU VAL GLY ASP VAL VAL ALA LEU LEU GLU ALA
SEQRES   8 B  328  ILE ALA LEU ASN GLU GLU LYS VAL PHE VAL VAL ALA HIS
SEQRES   9 B  328  ASP LEU GLY ALA TYR ILE ALA TRP HIS LEU CYS LEU PHE
SEQRES  10 B  328  ARG PRO ASP LYS VAL LYS ALA LEU VAL ASN LEU SER VAL
SEQRES  11 B  328  HIS PHE SER LYS ARG ASN PRO LYS MET ASN LYS VAL GLU
SEQRES  12 B  328  GLY LEU LYS ALA ILE TYR GLY GLU ASP HIS TYR VAL SER
SEQRES  13 B  328  ARG PHE GLN VAL PRO GLY GLU ILE GLU ALA GLU PHE ALA
SEQRES  14 B  328  PRO ILE GLY ALA LYS SER VAL LEU LYS LYS ILE LEU THR
SEQRES  15 B  328  TYR ARG ASP PRO ALA PRO PHE TYR PHE PRO LYS GLY LYS
SEQRES  16 B  328  GLY LEU GLU ALA ILE PRO ASP ALA PRO VAL ALA LEU SER
SEQRES  17 B  328  SER TRP LEU SER GLU GLU GLU LEU ASP TYR TYR ALA ASN
SEQRES  18 B  328  LYS PHE GLU GLN THR GLY PHE THR GLY ALA VAL ASN TYR
SEQRES  19 B  328  TYR ARG ALA LEU PRO ILE ASN TRP GLU LEU THR ALA PRO
SEQRES  20 B  328  TRP THR GLY ALA GLN VAL LYS VAL PRO THR LYS PHE ILE
SEQRES  21 B  328  VAL GLY GLU PHE ASP LEU VAL TYR HIS ILE PRO GLY ALA
SEQRES  22 B  328  LYS GLU TYR ILE HIS ASN GLY GLY PHE LYS LYS ASP VAL
SEQRES  23 B  328  PRO LEU LEU GLU GLU VAL VAL VAL LEU GLU GLY ALA ALA
SEQRES  24 B  328  HIS PHE VAL SER GLN GLU ARG PRO HIS GLU ILE SER LYS
SEQRES  25 B  328  HIS ILE TYR ASP PHE ILE GLN LYS PHE THR SER HIS HIS
SEQRES  26 B  328  HIS HIS HIS
HET    DIO  A1322       6
HET    DIO  B1322       6
HETNAM     DIO 1,4-DIETHYLENE DIOXIDE
FORMUL   3  DIO    2(C4 H8 O2)
FORMUL   4  HOH   *642(H2 O)
HELIX    1   1 LEU A   36  SER A   39  5                                   4
HELIX    2   2 TRP A   40  ARG A   50  1                                  11
HELIX    3   3 ASP A   72  PHE A   76  5                                   5
HELIX    4   4 SER A   77  ALA A   93  1                                  17
HELIX    5   5 ASP A  105  ARG A  118  1                                  14
HELIX    6   6 ASN A  140  GLY A  150  1                                  11
HELIX    7   7 HIS A  153  PHE A  158  1                                   6
HELIX    8   8 GLY A  162  ALA A  169  1                                   8
HELIX    9   9 GLY A  172  THR A  182  1                                  11
HELIX   10  10 PRO A  204  SER A  209  5                                   6
HELIX   11  11 SER A  212  GLY A  227  1                                  16
HELIX   12  12 PHE A  228  ALA A  237  1                                  10
HELIX   13  13 ALA A  237  THR A  245  1                                   9
HELIX   14  14 ALA A  246  THR A  249  5                                   4
HELIX   15  15 ASP A  265  ILE A  270  5                                   6
HELIX   16  16 GLY A  272  ASN A  279  1                                   8
HELIX   17  17 GLY A  280  VAL A  286  1                                   7
HELIX   18  18 PHE A  301  ARG A  306  1                                   6
HELIX   19  19 ARG A  306  GLN A  319  1                                  14
HELIX   20  20 LEU B   36  SER B   39  5                                   4
HELIX   21  21 TRP B   40  ARG B   50  1                                  11
HELIX   22  22 ASP B   72  PHE B   76  5                                   5
HELIX   23  23 SER B   77  ALA B   93  1                                  17
HELIX   24  24 ASP B  105  ARG B  118  1                                  14
HELIX   25  25 ASN B  140  GLY B  150  1                                  11
HELIX   26  26 HIS B  153  PHE B  158  1                                   6
HELIX   27  27 GLY B  162  ALA B  169  1                                   8
HELIX   28  28 GLY B  172  THR B  182  1                                  11
HELIX   29  29 PRO B  204  SER B  209  5                                   6
HELIX   30  30 SER B  212  GLY B  227  1                                  16
HELIX   31  31 PHE B  228  THR B  245  1                                  18
HELIX   32  32 ALA B  246  THR B  249  5                                   4
HELIX   33  33 ASP B  265  ILE B  270  5                                   6
HELIX   34  34 GLY B  272  ASN B  279  1                                   8
HELIX   35  35 GLY B  280  VAL B  286  1                                   7
HELIX   36  36 PHE B  301  ARG B  306  1                                   6
HELIX   37  37 ARG B  306  LYS B  320  1                                  15
SHEET    1  AA 8 GLU A   5  VAL A  11  0
SHEET    2  AA 8 LEU A  14  LEU A  21 -1  O  LEU A  14   N  VAL A  11
SHEET    3  AA 8 ARG A  53  PRO A  57 -1  O  ALA A  54   N  LEU A  21
SHEET    4  AA 8 THR A  26  ILE A  30  1  O  ILE A  27   N  VAL A  55
SHEET    5  AA 8 VAL A  99  HIS A 104  1  O  PHE A 100   N  LEU A  28
SHEET    6  AA 8 VAL A 122  LEU A 128  1  N  LYS A 123   O  VAL A  99
SHEET    7  AA 8 THR A 257  GLY A 262  1  O  LYS A 258   N  ASN A 127
SHEET    8  AA 8 VAL A 293  LEU A 295  1  O  VAL A 293   N  VAL A 261
SHEET    1  BA 8 GLU B   5  VAL B  11  0
SHEET    2  BA 8 LEU B  14  LEU B  21 -1  O  LEU B  14   N  VAL B  11
SHEET    3  BA 8 ARG B  53  PRO B  57 -1  O  ALA B  54   N  LEU B  21
SHEET    4  BA 8 THR B  26  ILE B  30  1  O  ILE B  27   N  VAL B  55
SHEET    5  BA 8 VAL B  99  HIS B 104  1  O  PHE B 100   N  LEU B  28
SHEET    6  BA 8 VAL B 122  LEU B 128  1  N  LYS B 123   O  VAL B  99
SHEET    7  BA 8 THR B 257  GLY B 262  1  O  LYS B 258   N  ASN B 127
SHEET    8  BA 8 VAL B 293  LEU B 295  1  O  VAL B 293   N  VAL B 261
CISPEP   1 PHE A   33    PRO A   34          0       -11.06
CISPEP   2 PHE B   33    PRO B   34          0        -9.82
SITE     1 AC1  5 PHE A  33  TYR A 154  PHE A 189  HIS A 300
SITE     2 AC1  5 HOH A2133
SITE     1 AC2  4 PHE B  33  TYR B 154  LEU B 266  HIS B 300
CRYST1   56.003   99.140  123.461  90.00  90.00  90.00 P 21 21 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      0.017856  0.000000  0.000000        0.00000
SCALE2      0.000000  0.010087  0.000000        0.00000
SCALE3      0.000000  0.000000  0.008100        0.00000
MTRIX1   1  0.704500  0.692500  0.155500      -74.64000    1
MTRIX2   1  0.665000 -0.567500 -0.485600      117.40000    1
MTRIX3   1 -0.248000  0.445500 -0.860300       20.09000    1
TER    2590      THR A 322
TER    5160      THR B 322
MASTER      334    0    2   37   16    0    3    9 5813    2   12   52
END