HEADER HYDROLASE 05-MAY-05 1ZL6
TITLE CRYSTAL STRUCTURE OF TYR350ALA MUTANT OF CLOSTRIDIUM BOTULINUM
TITLE 2 NEUROTOXIN E CATALYTIC DOMAIN
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: BOTULINUM NEUROTOXIN TYPE E;
COMPND 3 CHAIN: A, B;
COMPND 4 FRAGMENT: CATALYTIC DOMAIN (RESIDUES 2-421);
COMPND 5 EC: 3.4.24.69;
COMPND 6 ENGINEERED: YES;
COMPND 7 MUTATION: YES
SOURCE MOL_ID: 1;
SOURCE 2 ORGANISM_SCIENTIFIC: CLOSTRIDIUM BOTULINUM;
SOURCE 3 ORGANISM_TAXID: 1491;
SOURCE 4 STRAIN: NCTC-11219;
SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI BL21(DE3);
SOURCE 6 EXPRESSION_SYSTEM_TAXID: 469008;
SOURCE 7 EXPRESSION_SYSTEM_STRAIN: BL21(DE3);
SOURCE 8 EXPRESSION_SYSTEM_VECTOR_TYPE: T7;
SOURCE 9 EXPRESSION_SYSTEM_PLASMID: PET-9C
KEYWDS BOTULINUM NEUROTOXIN E, CATALYTIC DOMAIN, LIGHT CHAIN, TYR350ALA
KEYWDS 2 MUTANT, HYDROLASE
EXPDTA X-RAY DIFFRACTION
AUTHOR R.AGARWAL,T.BINZ,S.SWAMINATHAN
REVDAT 5 23-AUG-23 1ZL6 1 REMARK
REVDAT 4 20-OCT-21 1ZL6 1 REMARK SEQADV LINK
REVDAT 3 11-OCT-17 1ZL6 1 REMARK
REVDAT 2 24-FEB-09 1ZL6 1 VERSN
REVDAT 1 28-JUN-05 1ZL6 0
JRNL AUTH R.AGARWAL,T.BINZ,S.SWAMINATHAN
JRNL TITL ANALYSIS OF ACTIVE SITE RESIDUES OF BOTULINUM NEUROTOXIN E
JRNL TITL 2 BY MUTATIONAL, FUNCTIONAL, AND STRUCTURAL STUDIES: GLU335GLN
JRNL TITL 3 IS AN APOENZYME.
JRNL REF BIOCHEMISTRY V. 44 8291 2005
JRNL REFN ISSN 0006-2960
JRNL PMID 15938619
JRNL DOI 10.1021/BI050253A
REMARK 2
REMARK 2 RESOLUTION. 2.40 ANGSTROMS.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : CNS 1.1
REMARK 3 AUTHORS : BRUNGER,ADAMS,CLORE,DELANO,GROS,GROSSE-
REMARK 3 : KUNSTLEVE,JIANG,KUSZEWSKI,NILGES,PANNU,
REMARK 3 : READ,RICE,SIMONSON,WARREN
REMARK 3
REMARK 3 REFINEMENT TARGET : ENGH & HUBER
REMARK 3
REMARK 3 DATA USED IN REFINEMENT.
REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.40
REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 46.54
REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.000
REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 225939.930
REMARK 3 DATA CUTOFF LOW (ABS(F)) : 0.0000
REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 92.8
REMARK 3 NUMBER OF REFLECTIONS : 39653
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 SET) : 0.235
REMARK 3 FREE R VALUE : 0.284
REMARK 3 FREE R VALUE TEST SET SIZE (%) : 3.000
REMARK 3 FREE R VALUE TEST SET COUNT : 1206
REMARK 3 ESTIMATED ERROR OF FREE R VALUE : 0.008
REMARK 3
REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN.
REMARK 3 TOTAL NUMBER OF BINS USED : 6
REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 2.40
REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.55
REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 80.70
REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 5463
REMARK 3 BIN R VALUE (WORKING SET) : 0.3610
REMARK 3 BIN FREE R VALUE : 0.4170
REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 3.30
REMARK 3 BIN FREE R VALUE TEST SET COUNT : 186
REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.031
REMARK 3
REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
REMARK 3 PROTEIN ATOMS : 6303
REMARK 3 NUCLEIC ACID ATOMS : 0
REMARK 3 HETEROGEN ATOMS : 7
REMARK 3 SOLVENT ATOMS : 207
REMARK 3
REMARK 3 B VALUES.
REMARK 3 FROM WILSON PLOT (A**2) : 32.00
REMARK 3 MEAN B VALUE (OVERALL, A**2) : 48.70
REMARK 3 OVERALL ANISOTROPIC B VALUE.
REMARK 3 B11 (A**2) : 9.54000
REMARK 3 B22 (A**2) : -15.02000
REMARK 3 B33 (A**2) : 5.49000
REMARK 3 B12 (A**2) : 0.00000
REMARK 3 B13 (A**2) : 0.00000
REMARK 3 B23 (A**2) : 0.00000
REMARK 3
REMARK 3 ESTIMATED COORDINATE ERROR.
REMARK 3 ESD FROM LUZZATI PLOT (A) : 0.34
REMARK 3 ESD FROM SIGMAA (A) : 0.46
REMARK 3 LOW RESOLUTION CUTOFF (A) : 5.00
REMARK 3
REMARK 3 CROSS-VALIDATED ESTIMATED COORDINATE ERROR.
REMARK 3 ESD FROM C-V LUZZATI PLOT (A) : 0.42
REMARK 3 ESD FROM C-V SIGMAA (A) : 0.53
REMARK 3
REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES.
REMARK 3 BOND LENGTHS (A) : 0.007
REMARK 3 BOND ANGLES (DEGREES) : 1.300
REMARK 3 DIHEDRAL ANGLES (DEGREES) : 22.90
REMARK 3 IMPROPER ANGLES (DEGREES) : 0.750
REMARK 3
REMARK 3 ISOTROPIC THERMAL MODEL : RESTRAINED
REMARK 3
REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA
REMARK 3 MAIN-CHAIN BOND (A**2) : 1.290 ; 1.500
REMARK 3 MAIN-CHAIN ANGLE (A**2) : 2.170 ; 2.000
REMARK 3 SIDE-CHAIN BOND (A**2) : 1.870 ; 2.000
REMARK 3 SIDE-CHAIN ANGLE (A**2) : 2.790 ; 2.500
REMARK 3
REMARK 3 BULK SOLVENT MODELING.
REMARK 3 METHOD USED : FLAT MODEL
REMARK 3 KSOL : 0.34
REMARK 3 BSOL : 34.39
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 : PROTEIN_REP.PARAM
REMARK 3 PARAMETER FILE 2 : ION.PARAM
REMARK 3 PARAMETER FILE 3 : WATER_REP.PARAM
REMARK 3 PARAMETER FILE 4 : NULL
REMARK 3 TOPOLOGY FILE 1 : PROTEIN.TOP
REMARK 3 TOPOLOGY FILE 2 : NULL
REMARK 3 TOPOLOGY FILE 3 : NULL
REMARK 3 TOPOLOGY FILE 4 : NULL
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS:
REMARK 3 IN GENERAL, THREE LOOP REGIONS (50-65, 188-200 AND 234-244) ARE
REMARK 3 EITHER
REMARK 3 DISORDERED OR HAVE VERY WEAK ELECTRON DENSITY IN BOTH CHAINS. IN
REMARK 3 THIS ENTRY,
REMARK 3 THE MISSING RESIDUES ARE DUE TO THE ABSENCE OF INTERPRETABLE
REMARK 3 ELECTRON
REMARK 3 DENSITY IN THOSE REGIONS.
REMARK 4
REMARK 4 1ZL6 COMPLIES WITH FORMAT V. 3.30, 13-JUL-11
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 11-MAY-05.
REMARK 100 THE DEPOSITION ID IS D_1000032846.
REMARK 200
REMARK 200 EXPERIMENTAL DETAILS
REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION
REMARK 200 DATE OF DATA COLLECTION : 04-AUG-04
REMARK 200 TEMPERATURE (KELVIN) : 100
REMARK 200 PH : 5.6
REMARK 200 NUMBER OF CRYSTALS USED : 1
REMARK 200
REMARK 200 SYNCHROTRON (Y/N) : Y
REMARK 200 RADIATION SOURCE : NSLS
REMARK 200 BEAMLINE : X25
REMARK 200 X-RAY GENERATOR MODEL : NULL
REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M
REMARK 200 WAVELENGTH OR RANGE (A) : 1.1
REMARK 200 MONOCHROMATOR : GRAPHITE
REMARK 200 OPTICS : MIRRORS
REMARK 200
REMARK 200 DETECTOR TYPE : CCD
REMARK 200 DETECTOR MANUFACTURER : ADSC QUANTUM 315
REMARK 200 INTENSITY-INTEGRATION SOFTWARE : CBASS
REMARK 200 DATA SCALING SOFTWARE : HKL-2000
REMARK 200
REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 42956
REMARK 200 RESOLUTION RANGE HIGH (A) : 2.400
REMARK 200 RESOLUTION RANGE LOW (A) : 50.000
REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 0.000
REMARK 200
REMARK 200 OVERALL.
REMARK 200 COMPLETENESS FOR RANGE (%) : 99.9
REMARK 200 DATA REDUNDANCY : 5.900
REMARK 200 R MERGE (I) : 0.08000
REMARK 200 R SYM (I) : NULL
REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : 9.0000
REMARK 200
REMARK 200 IN THE HIGHEST RESOLUTION SHELL.
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.40
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.49
REMARK 200 COMPLETENESS FOR SHELL (%) : 99.4
REMARK 200 DATA REDUNDANCY IN SHELL : 4.70
REMARK 200 R MERGE FOR SHELL (I) : 0.44000
REMARK 200 R SYM FOR SHELL (I) : NULL
REMARK 200 <I/SIGMA(I)> FOR SHELL : 3.000
REMARK 200
REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH
REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: FOURIER SYNTHESIS
REMARK 200 SOFTWARE USED: REFINED WITH THE MODEL 1T3A
REMARK 200 STARTING MODEL: PDB ENTRY 1T3A
REMARK 200
REMARK 200 REMARK: NULL
REMARK 280
REMARK 280 CRYSTAL
REMARK 280 SOLVENT CONTENT, VS (%): 48.00
REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.67
REMARK 280
REMARK 280 CRYSTALLIZATION CONDITIONS: AMMONIUM SULFATE, LITHIUM SULFATE,
REMARK 280 SODIUM CITRATE, PH 5.6, VAPOR DIFFUSION, SITTING DROP,
REMARK 280 TEMPERATURE 298K
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY
REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: P 21 21 2
REMARK 290
REMARK 290 SYMOP SYMMETRY
REMARK 290 NNNMMM OPERATOR
REMARK 290 1555 X,Y,Z
REMARK 290 2555 -X,-Y,Z
REMARK 290 3555 -X+1/2,Y+1/2,-Z
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 0.00000
REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000
REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 0.00000
REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 44.40000
REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 72.47900
REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 0.00000
REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 44.40000
REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 72.47900
REMARK 290 SMTRY3 4 0.000000 0.000000 -1.000000 0.00000
REMARK 290
REMARK 290 REMARK: NULL
REMARK 300
REMARK 300 BIOMOLECULE: 1
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: DIMERIC
REMARK 350 APPLY THE FOLLOWING TO CHAINS: A, 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 GLN A 234
REMARK 465 LYS A 235
REMARK 465 GLN A 236
REMARK 465 ASN A 237
REMARK 465 PRO A 238
REMARK 465 LEU A 239
REMARK 465 ILE A 240
REMARK 465 THR A 241
REMARK 465 ASN A 242
REMARK 465 ILE A 243
REMARK 465 ARG A 244
REMARK 465 LYS A 412
REMARK 465 ASN A 413
REMARK 465 ILE A 414
REMARK 465 VAL A 415
REMARK 465 SER A 416
REMARK 465 VAL A 417
REMARK 465 LYS A 418
REMARK 465 GLY A 419
REMARK 465 ILE A 420
REMARK 465 HIS B 55
REMARK 465 PRO B 56
REMARK 465 PRO B 57
REMARK 465 THR B 58
REMARK 465 SER B 59
REMARK 465 LEU B 60
REMARK 465 LYS B 61
REMARK 465 ASN B 62
REMARK 465 GLY B 63
REMARK 465 ASP B 64
REMARK 465 SER B 65
REMARK 465 GLN B 234
REMARK 465 LYS B 235
REMARK 465 GLN B 236
REMARK 465 ASN B 237
REMARK 465 PRO B 238
REMARK 465 LEU B 239
REMARK 465 ILE B 240
REMARK 465 THR B 241
REMARK 465 ASN B 242
REMARK 465 ILE B 243
REMARK 465 ARG B 244
REMARK 465 PHE B 410
REMARK 465 CYS B 411
REMARK 465 LYS B 412
REMARK 465 ASN B 413
REMARK 465 ILE B 414
REMARK 465 VAL B 415
REMARK 465 SER B 416
REMARK 465 VAL B 417
REMARK 465 LYS B 418
REMARK 465 GLY B 419
REMARK 465 ILE B 420
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 ARG A 15 -72.18 -116.21
REMARK 500 LEU A 18 -168.99 -162.44
REMARK 500 LYS A 36 110.09 -35.12
REMARK 500 PRO A 51 -33.39 -39.90
REMARK 500 ASP A 69 100.82 -166.10
REMARK 500 ALA A 109 48.64 -89.18
REMARK 500 ASN A 247 153.75 -47.94
REMARK 500 GLN A 289 150.34 -42.02
REMARK 500 ASN A 361 105.49 -21.46
REMARK 500 ASN A 377 -126.43 45.35
REMARK 500 THR A 399 129.63 -37.44
REMARK 500 ARG A 409 88.93 65.54
REMARK 500 ASP B 14 11.91 57.58
REMARK 500 ARG B 15 -62.66 -129.26
REMARK 500 LYS B 36 113.37 -37.67
REMARK 500 PRO B 42 64.64 -68.20
REMARK 500 ASN B 45 96.73 -63.22
REMARK 500 ASP B 53 -34.28 -38.15
REMARK 500 ASP B 69 89.47 178.20
REMARK 500 ASN B 95 -7.65 -54.50
REMARK 500 ALA B 109 30.85 -87.01
REMARK 500 ASP B 141 -151.32 -150.49
REMARK 500 MET B 198 41.22 15.93
REMARK 500 THR B 227 -3.95 -54.39
REMARK 500 ASN B 361 107.42 -25.42
REMARK 500 ASN B 375 80.19 -63.09
REMARK 500 ASN B 377 -117.29 47.51
REMARK 500 ASN B 382 37.96 38.30
REMARK 500 ASN B 391 74.07 -157.47
REMARK 500
REMARK 500 REMARK: NULL
REMARK 620
REMARK 620 METAL COORDINATION
REMARK 620 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER;
REMARK 620 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE):
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 ZN A 422 ZN
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 HIS A 211 NE2
REMARK 620 2 HIS A 215 NE2 95.5
REMARK 620 3 GLU A 250 OE1 131.9 96.5
REMARK 620 4 SO4 A 500 O3 101.4 92.3 124.4
REMARK 620 5 SO4 A 500 O1 137.9 119.2 70.8 57.3
REMARK 620 N 1 2 3 4
REMARK 620
REMARK 620 COORDINATION ANGLES FOR: M RES CSSEQI METAL
REMARK 620 ZN B 822 ZN
REMARK 620 N RES CSSEQI ATOM
REMARK 620 1 HIS B 211 NE2
REMARK 620 2 HIS B 215 NE2 96.9
REMARK 620 3 GLU B 250 OE2 140.7 61.5
REMARK 620 4 GLU B 250 OE1 90.9 60.6 50.2
REMARK 620 N 1 2 3
REMARK 800
REMARK 800 SITE
REMARK 800 SITE_IDENTIFIER: AC1
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE ZN A 422
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC2
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE ZN B 822
REMARK 800
REMARK 800 SITE_IDENTIFIER: AC3
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 500
REMARK 900
REMARK 900 RELATED ENTRIES
REMARK 900 RELATED ID: 1T3A RELATED DB: PDB
REMARK 900 CRYSTAL STRUCTURE OF CLOSTRIDIUM BOTULINUM NEUROTOXIN E CATALYTIC
REMARK 900 DOMAIN
REMARK 900 RELATED ID: 1T3C RELATED DB: PDB
REMARK 900 CRYSTAL STRUCTURE OF CLOSTRIDIUM BOTULINUM NEUROTOXIN E GLU212GLN
REMARK 900 MUTANT OF CATALYTIC DOMAIN
REMARK 900 RELATED ID: 1ZKW RELATED DB: PDB
REMARK 900 CRYSTAL STRUCTURE OF ARG347ALA MUTANT OF CLOSTRIDIUM BOTULINUM
REMARK 900 NEUROTOXIN E CATALYTIC DOMAIN
REMARK 900 RELATED ID: 1ZKX RELATED DB: PDB
REMARK 900 CRYSTAL STRUCTURE OF GLU158ALA/THR159ALA/ASN160ALA-TRIPLE MUTANT OF
REMARK 900 MUTANT OF CLOSTRIDIUM BOTULINUM NEUROTOXIN E CATALYTIC DOMAIN
REMARK 900 RELATED ID: 1ZL5 RELATED DB: PDB
REMARK 900 CRYSTAL STRUCTURE OF GLU335GLN MUTANT OF CLOSTRIDIUM BOTULINUM
REMARK 900 NEUROTOXIN E CATALYTIC DOMAIN
DBREF 1ZL6 A 1 420 UNP Q00496 BXE_CLOBO 2 421
DBREF 1ZL6 B 1 420 UNP Q00496 BXE_CLOBO 2 421
SEQADV 1ZL6 ALA A 350 UNP Q00496 TYR 351 ENGINEERED MUTATION
SEQADV 1ZL6 ALA B 350 UNP Q00496 TYR 351 ENGINEERED MUTATION
SEQRES 1 A 420 PRO LYS ILE ASN SER PHE ASN TYR ASN ASP PRO VAL ASN
SEQRES 2 A 420 ASP ARG THR ILE LEU TYR ILE LYS PRO GLY GLY CYS GLN
SEQRES 3 A 420 GLU PHE TYR LYS SER PHE ASN ILE MET LYS ASN ILE TRP
SEQRES 4 A 420 ILE ILE PRO GLU ARG ASN VAL ILE GLY THR THR PRO GLN
SEQRES 5 A 420 ASP PHE HIS PRO PRO THR SER LEU LYS ASN GLY ASP SER
SEQRES 6 A 420 SER TYR TYR ASP PRO ASN TYR LEU GLN SER ASP GLU GLU
SEQRES 7 A 420 LYS ASP ARG PHE LEU LYS ILE VAL THR LYS ILE PHE ASN
SEQRES 8 A 420 ARG ILE ASN ASN ASN LEU SER GLY GLY ILE LEU LEU GLU
SEQRES 9 A 420 GLU LEU SER LYS ALA ASN PRO TYR LEU GLY ASN ASP ASN
SEQRES 10 A 420 THR PRO ASP ASN GLN PHE HIS ILE GLY ASP ALA SER ALA
SEQRES 11 A 420 VAL GLU ILE LYS PHE SER ASN GLY SER GLN ASP ILE LEU
SEQRES 12 A 420 LEU PRO ASN VAL ILE ILE MET GLY ALA GLU PRO ASP LEU
SEQRES 13 A 420 PHE GLU THR ASN SER SER ASN ILE SER LEU ARG ASN ASN
SEQRES 14 A 420 TYR MET PRO SER ASN HIS GLY PHE GLY SER ILE ALA ILE
SEQRES 15 A 420 VAL THR PHE SER PRO GLU TYR SER PHE ARG PHE ASN ASP
SEQRES 16 A 420 ASN SER MET ASN GLU PHE ILE GLN ASP PRO ALA LEU THR
SEQRES 17 A 420 LEU MET HIS GLU LEU ILE HIS SER LEU HIS GLY LEU TYR
SEQRES 18 A 420 GLY ALA LYS GLY ILE THR THR LYS TYR THR ILE THR GLN
SEQRES 19 A 420 LYS GLN ASN PRO LEU ILE THR ASN ILE ARG GLY THR ASN
SEQRES 20 A 420 ILE GLU GLU PHE LEU THR PHE GLY GLY THR ASP LEU ASN
SEQRES 21 A 420 ILE ILE THR SER ALA GLN SER ASN ASP ILE TYR THR ASN
SEQRES 22 A 420 LEU LEU ALA ASP TYR LYS LYS ILE ALA SER LYS LEU SER
SEQRES 23 A 420 LYS VAL GLN VAL SER ASN PRO LEU LEU ASN PRO TYR LYS
SEQRES 24 A 420 ASP VAL PHE GLU ALA LYS TYR GLY LEU ASP LYS ASP ALA
SEQRES 25 A 420 SER GLY ILE TYR SER VAL ASN ILE ASN LYS PHE ASN ASP
SEQRES 26 A 420 ILE PHE LYS LYS LEU TYR SER PHE THR GLU PHE ASP LEU
SEQRES 27 A 420 ALA THR LYS PHE GLN VAL LYS CYS ARG GLN THR ALA ILE
SEQRES 28 A 420 GLY GLN TYR LYS TYR PHE LYS LEU SER ASN LEU LEU ASN
SEQRES 29 A 420 ASP SER ILE TYR ASN ILE SER GLU GLY TYR ASN ILE ASN
SEQRES 30 A 420 ASN LEU LYS VAL ASN PHE ARG GLY GLN ASN ALA ASN LEU
SEQRES 31 A 420 ASN PRO ARG ILE ILE THR PRO ILE THR GLY ARG GLY LEU
SEQRES 32 A 420 VAL LYS LYS ILE ILE ARG PHE CYS LYS ASN ILE VAL SER
SEQRES 33 A 420 VAL LYS GLY ILE
SEQRES 1 B 420 PRO LYS ILE ASN SER PHE ASN TYR ASN ASP PRO VAL ASN
SEQRES 2 B 420 ASP ARG THR ILE LEU TYR ILE LYS PRO GLY GLY CYS GLN
SEQRES 3 B 420 GLU PHE TYR LYS SER PHE ASN ILE MET LYS ASN ILE TRP
SEQRES 4 B 420 ILE ILE PRO GLU ARG ASN VAL ILE GLY THR THR PRO GLN
SEQRES 5 B 420 ASP PHE HIS PRO PRO THR SER LEU LYS ASN GLY ASP SER
SEQRES 6 B 420 SER TYR TYR ASP PRO ASN TYR LEU GLN SER ASP GLU GLU
SEQRES 7 B 420 LYS ASP ARG PHE LEU LYS ILE VAL THR LYS ILE PHE ASN
SEQRES 8 B 420 ARG ILE ASN ASN ASN LEU SER GLY GLY ILE LEU LEU GLU
SEQRES 9 B 420 GLU LEU SER LYS ALA ASN PRO TYR LEU GLY ASN ASP ASN
SEQRES 10 B 420 THR PRO ASP ASN GLN PHE HIS ILE GLY ASP ALA SER ALA
SEQRES 11 B 420 VAL GLU ILE LYS PHE SER ASN GLY SER GLN ASP ILE LEU
SEQRES 12 B 420 LEU PRO ASN VAL ILE ILE MET GLY ALA GLU PRO ASP LEU
SEQRES 13 B 420 PHE GLU THR ASN SER SER ASN ILE SER LEU ARG ASN ASN
SEQRES 14 B 420 TYR MET PRO SER ASN HIS GLY PHE GLY SER ILE ALA ILE
SEQRES 15 B 420 VAL THR PHE SER PRO GLU TYR SER PHE ARG PHE ASN ASP
SEQRES 16 B 420 ASN SER MET ASN GLU PHE ILE GLN ASP PRO ALA LEU THR
SEQRES 17 B 420 LEU MET HIS GLU LEU ILE HIS SER LEU HIS GLY LEU TYR
SEQRES 18 B 420 GLY ALA LYS GLY ILE THR THR LYS TYR THR ILE THR GLN
SEQRES 19 B 420 LYS GLN ASN PRO LEU ILE THR ASN ILE ARG GLY THR ASN
SEQRES 20 B 420 ILE GLU GLU PHE LEU THR PHE GLY GLY THR ASP LEU ASN
SEQRES 21 B 420 ILE ILE THR SER ALA GLN SER ASN ASP ILE TYR THR ASN
SEQRES 22 B 420 LEU LEU ALA ASP TYR LYS LYS ILE ALA SER LYS LEU SER
SEQRES 23 B 420 LYS VAL GLN VAL SER ASN PRO LEU LEU ASN PRO TYR LYS
SEQRES 24 B 420 ASP VAL PHE GLU ALA LYS TYR GLY LEU ASP LYS ASP ALA
SEQRES 25 B 420 SER GLY ILE TYR SER VAL ASN ILE ASN LYS PHE ASN ASP
SEQRES 26 B 420 ILE PHE LYS LYS LEU TYR SER PHE THR GLU PHE ASP LEU
SEQRES 27 B 420 ALA THR LYS PHE GLN VAL LYS CYS ARG GLN THR ALA ILE
SEQRES 28 B 420 GLY GLN TYR LYS TYR PHE LYS LEU SER ASN LEU LEU ASN
SEQRES 29 B 420 ASP SER ILE TYR ASN ILE SER GLU GLY TYR ASN ILE ASN
SEQRES 30 B 420 ASN LEU LYS VAL ASN PHE ARG GLY GLN ASN ALA ASN LEU
SEQRES 31 B 420 ASN PRO ARG ILE ILE THR PRO ILE THR GLY ARG GLY LEU
SEQRES 32 B 420 VAL LYS LYS ILE ILE ARG PHE CYS LYS ASN ILE VAL SER
SEQRES 33 B 420 VAL LYS GLY ILE
HET ZN A 422 1
HET SO4 A 500 5
HET ZN B 822 1
HETNAM ZN ZINC ION
HETNAM SO4 SULFATE ION
FORMUL 3 ZN 2(ZN 2+)
FORMUL 4 SO4 O4 S 2-
FORMUL 6 HOH *207(H2 O)
HELIX 1 1 THR A 50 HIS A 55 5 6
HELIX 2 2 SER A 75 ASN A 95 1 21
HELIX 3 3 ASN A 96 ALA A 109 1 14
HELIX 4 4 LEU A 166 TYR A 170 5 5
HELIX 5 5 MET A 171 HIS A 175 5 5
HELIX 6 6 ASP A 204 TYR A 221 1 18
HELIX 7 7 ASN A 247 GLY A 255 1 9
HELIX 8 8 GLY A 256 ILE A 262 5 7
HELIX 9 9 THR A 263 LYS A 287 1 25
HELIX 10 10 ASN A 292 LEU A 294 5 3
HELIX 11 11 LEU A 295 TYR A 306 1 12
HELIX 12 12 ASN A 319 TYR A 331 1 13
HELIX 13 13 THR A 334 GLN A 343 1 10
HELIX 14 14 ILE A 376 ASN A 387 5 12
HELIX 15 15 ASN A 391 ARG A 393 5 3
HELIX 16 16 GLY A 402 ILE A 408 1 7
HELIX 17 17 THR B 50 PHE B 54 5 5
HELIX 18 18 SER B 75 ASN B 95 1 21
HELIX 19 19 ASN B 96 LYS B 108 1 13
HELIX 20 20 LEU B 166 TYR B 170 5 5
HELIX 21 21 MET B 171 HIS B 175 5 5
HELIX 22 22 ASP B 204 TYR B 221 1 18
HELIX 23 23 ASN B 247 GLY B 255 1 9
HELIX 24 24 GLY B 256 LEU B 259 5 4
HELIX 25 25 THR B 263 VAL B 288 1 26
HELIX 26 26 ASN B 292 LEU B 295 5 4
HELIX 27 27 ASN B 296 TYR B 306 1 11
HELIX 28 28 ASN B 319 TYR B 331 1 13
HELIX 29 29 THR B 334 GLN B 343 1 10
HELIX 30 30 ILE B 376 ASN B 387 5 12
HELIX 31 31 ASN B 391 ILE B 395 5 5
HELIX 32 32 GLY B 400 ILE B 408 1 9
SHEET 1 A 8 GLN A 140 LEU A 143 0
SHEET 2 A 8 VAL A 131 LYS A 134 -1 N ILE A 133 O ASP A 141
SHEET 3 A 8 ILE A 17 LYS A 21 -1 N LYS A 21 O GLU A 132
SHEET 4 A 8 TYR A 29 MET A 35 -1 O SER A 31 N LEU A 18
SHEET 5 A 8 ILE A 38 ARG A 44 -1 O ILE A 38 N MET A 35
SHEET 6 A 8 VAL A 147 GLY A 151 1 O ILE A 149 N TRP A 39
SHEET 7 A 8 ALA A 181 THR A 184 1 O VAL A 183 N MET A 150
SHEET 8 A 8 ASN A 160 ASN A 163 -1 N SER A 162 O ILE A 182
SHEET 1 B 4 GLU A 200 ILE A 202 0
SHEET 2 B 4 TYR A 189 ASN A 194 -1 N PHE A 193 O PHE A 201
SHEET 3 B 4 TYR A 356 LEU A 359 -1 O PHE A 357 N SER A 190
SHEET 4 B 4 ILE A 395 THR A 396 -1 O THR A 396 N LYS A 358
SHEET 1 C 2 LEU A 308 LYS A 310 0
SHEET 2 C 2 TYR A 316 VAL A 318 -1 O SER A 317 N ASP A 309
SHEET 1 D 8 GLN B 140 LEU B 143 0
SHEET 2 D 8 VAL B 131 LYS B 134 -1 N VAL B 131 O LEU B 143
SHEET 3 D 8 ILE B 17 LYS B 21 -1 N LYS B 21 O GLU B 132
SHEET 4 D 8 TYR B 29 MET B 35 -1 O SER B 31 N LEU B 18
SHEET 5 D 8 ILE B 38 ARG B 44 -1 O ILE B 40 N PHE B 32
SHEET 6 D 8 VAL B 147 GLY B 151 1 O ILE B 149 N TRP B 39
SHEET 7 D 8 ALA B 181 THR B 184 1 O VAL B 183 N MET B 150
SHEET 8 D 8 ASN B 160 ASN B 163 -1 N SER B 162 O ILE B 182
SHEET 1 E 3 GLU B 200 ILE B 202 0
SHEET 2 E 3 TYR B 189 ASN B 194 -1 N PHE B 193 O PHE B 201
SHEET 3 E 3 LYS B 355 LYS B 358 -1 O PHE B 357 N SER B 190
SHEET 1 F 2 LEU B 308 LYS B 310 0
SHEET 2 F 2 TYR B 316 VAL B 318 -1 O SER B 317 N ASP B 309
LINK NE2 HIS A 211 ZN ZN A 422 1555 1555 2.05
LINK NE2 HIS A 215 ZN ZN A 422 1555 1555 2.10
LINK OE1 GLU A 250 ZN ZN A 422 1555 1555 2.49
LINK ZN ZN A 422 O3 SO4 A 500 1555 1555 2.61
LINK ZN ZN A 422 O1 SO4 A 500 1555 1555 2.41
LINK NE2 HIS B 211 ZN ZN B 822 1555 1555 2.09
LINK NE2 HIS B 215 ZN ZN B 822 1555 1555 2.05
LINK OE2 GLU B 250 ZN ZN B 822 1555 1555 2.74
LINK OE1 GLU B 250 ZN ZN B 822 1555 1555 2.40
SITE 1 AC1 4 HIS A 211 HIS A 215 GLU A 250 SO4 A 500
SITE 1 AC2 3 HIS B 211 HIS B 215 GLU B 250
SITE 1 AC3 5 HIS A 211 GLU A 212 HIS A 215 GLU A 250
SITE 2 AC3 5 ZN A 422
CRYST1 88.800 144.958 83.185 90.00 90.00 90.00 P 21 21 2 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 0.011261 0.000000 0.000000 0.00000
SCALE2 0.000000 0.006899 0.000000 0.00000
SCALE3 0.000000 0.000000 0.012021 0.00000
(ATOM LINES ARE NOT SHOWN.)
END
