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HEADER HYDROLASE 17-JUN-04 1TQH
TITLE COVALENT REACTION INTERMEDIATE REVEALED IN CRYSTAL
TITLE 2 STRUCTURE OF THE GEOBACILLUS STEAROTHERMOPHILUS
TITLE 3 CARBOXYLESTERASE EST30
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: CARBOXYLESTERASE PRECURSOR;
COMPND 3 CHAIN: A;
COMPND 4 EC: 3.1.1.1;
COMPND 5 ENGINEERED: YES
SOURCE MOL_ID: 1;
SOURCE 2 ORGANISM_SCIENTIFIC: BACILLUS STEAROTHERMOPHILUS;
SOURCE 3 ORGANISM_COMMON: BACTERIA;
SOURCE 4 EXPRESSION_SYSTEM: ESCHERICHIA COLI;
SOURCE 5 EXPRESSION_SYSTEM_COMMON: BACTERIA
KEYWDS CARBOXYLESTERASE, TETRAHEDRAL INTERMEDIATE, ALPHA/BETA
KEYWDS 2 HYDROLASE
EXPDTA X-RAY DIFFRACTION
AUTHOR P.LIU,Y.F.WANG,H.E.EWIS,A.T.ABDELAL,C.D.LU,R.W.HARRISON,
AUTHOR 2 I.T.WEBER
REVDAT 1 28-SEP-04 1TQH 0
JRNL AUTH P.LIU,Y.F.WANG,H.E.EWIS,A.T.ABDELAL,C.D.LU,
JRNL AUTH 2 R.W.HARRISON,I.T.WEBER
JRNL TITL COVALENT REACTION INTERMEDIATE REVEALED IN CRYSTAL
JRNL TITL 2 STRUCTURE OF THE GEOBACILLUS STEAROTHERMOPHILUS
JRNL TITL 3 CARBOXYLESTERASE EST30.
JRNL REF J.MOL.BIOL. V. 342 551 2004
JRNL REFN ASTM JMOBAK UK ISSN 0022-2836
REMARK 1
REMARK 2
REMARK 2 RESOLUTION. 1.63 ANGSTROMS.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : SHELXL97
REMARK 3 AUTHORS : G.M.SHELDRICK
REMARK 3
REMARK 3 DATA USED IN REFINEMENT.
REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.63
REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 10.00
REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.000
REMARK 3 COMPLETENESS FOR RANGE (%) : 96.7
REMARK 3 CROSS-VALIDATION METHOD : FREE R
REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM
REMARK 3
REMARK 3 FIT TO DATA USED IN REFINEMENT (NO CUTOFF).
REMARK 3 R VALUE (WORKING + TEST SET, NO CUTOFF) : 0.171
REMARK 3 R VALUE (WORKING SET, NO CUTOFF) : 0.168
REMARK 3 FREE R VALUE (NO CUTOFF) : 0.228
REMARK 3 FREE R VALUE TEST SET SIZE (%, NO CUTOFF) : 5.000
REMARK 3 FREE R VALUE TEST SET COUNT (NO CUTOFF) : 1768
REMARK 3 TOTAL NUMBER OF REFLECTIONS (NO CUTOFF) : 35376
REMARK 3
REMARK 3 FIT/AGREEMENT OF MODEL FOR DATA WITH F>4SIG(F).
REMARK 3 R VALUE (WORKING + TEST SET, F>4SIG(F)) : 0.164
REMARK 3 R VALUE (WORKING SET, F>4SIG(F)) : 0.161
REMARK 3 FREE R VALUE (F>4SIG(F)) : 0.220
REMARK 3 FREE R VALUE TEST SET SIZE (%, F>4SIG(F)) : 5.000
REMARK 3 FREE R VALUE TEST SET COUNT (F>4SIG(F)) : 1581
REMARK 3 TOTAL NUMBER OF REFLECTIONS (F>4SIG(F)) : 31675
REMARK 3
REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
REMARK 3 PROTEIN ATOMS : 1998
REMARK 3 NUCLEIC ACID ATOMS : 0
REMARK 3 HETEROGEN ATOMS : 17
REMARK 3 SOLVENT ATOMS : 153
REMARK 3
REMARK 3 MODEL REFINEMENT.
REMARK 3 OCCUPANCY SUM OF NON-HYDROGEN ATOMS : 2115.00
REMARK 3 OCCUPANCY SUM OF HYDROGEN ATOMS : 0.00
REMARK 3 NUMBER OF DISCRETELY DISORDERED RESIDUES : 9
REMARK 3 NUMBER OF LEAST-SQUARES PARAMETERS : 19536
REMARK 3 NUMBER OF RESTRAINTS : 24598
REMARK 3
REMARK 3 RMS DEVIATIONS FROM RESTRAINT TARGET VALUES.
REMARK 3 BOND LENGTHS (A) : 0.008
REMARK 3 ANGLE DISTANCES (A) : 0.024
REMARK 3 SIMILAR DISTANCES (NO TARGET VALUES) (A) : 0.000
REMARK 3 DISTANCES FROM RESTRAINT PLANES (A) : 0.025
REMARK 3 ZERO CHIRAL VOLUMES (A**3) : 0.038
REMARK 3 NON-ZERO CHIRAL VOLUMES (A**3) : 0.051
REMARK 3 ANTI-BUMPING DISTANCE RESTRAINTS (A) : 0.011
REMARK 3 RIGID-BOND ADP COMPONENTS (A**2) : 0.002
REMARK 3 SIMILAR ADP COMPONENTS (A**2) : 0.038
REMARK 3 APPROXIMATELY ISOTROPIC ADPS (A**2) : 0.087
REMARK 3
REMARK 3 BULK SOLVENT MODELING.
REMARK 3 METHOD USED: NULL
REMARK 3
REMARK 3 STEREOCHEMISTRY TARGET VALUES : ENGH AND HUBER
REMARK 3 SPECIAL CASE: NULL
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS: NULL
REMARK 4
REMARK 4 1TQH COMPLIES WITH FORMAT V. 2.3, 09-JULY-1998
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 06-JUL-2004.
REMARK 100 THE RCSB ID CODE IS RCSB022838.
REMARK 200
REMARK 200 EXPERIMENTAL DETAILS
REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION
REMARK 200 DATE OF DATA COLLECTION : 22-OCT-2003; 06-MAR-2003
REMARK 200 TEMPERATURE (KELVIN) : 95; 95
REMARK 200 PH : 7.50
REMARK 200 NUMBER OF CRYSTALS USED : 2
REMARK 200
REMARK 200 SYNCHROTRON (Y/N) : Y; Y
REMARK 200 RADIATION SOURCE : SER-CAT22; SER-CAT22
REMARK 200 BEAMLINE : NULL
REMARK 200 X-RAY GENERATOR MODEL : NULL
REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M; M
REMARK 200 WAVELENGTH OR RANGE (A) : 0.99997; 0.97920, 0.97933,
REMARK 200 0.971602, 0.987072
REMARK 200 MONOCHROMATOR : SI; SI
REMARK 200 OPTICS : NULL
REMARK 200
REMARK 200 DETECTOR TYPE : MAR CCD 225; MAR CCD 165
REMARK 200 DETECTOR MANUFACTURER : MAR CCD; MAR CCD
REMARK 200 INTENSITY-INTEGRATION SOFTWARE : MARCCD
REMARK 200 DATA SCALING SOFTWARE : HKL2000
REMARK 200
REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 36452
REMARK 200 RESOLUTION RANGE HIGH (A) : 1.630
REMARK 200 RESOLUTION RANGE LOW (A) : 50.000
REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL
REMARK 200
REMARK 200 OVERALL.
REMARK 200 COMPLETENESS FOR RANGE (%) : 97.5
REMARK 200 DATA REDUNDANCY : 6.200
REMARK 200 R MERGE (I) : 0.15500
REMARK 200 R SYM (I) : NULL
REMARK 200 FOR THE DATA SET : NULL
REMARK 200
REMARK 200 IN THE HIGHEST RESOLUTION SHELL.
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.63
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.69
REMARK 200 COMPLETENESS FOR SHELL (%) : 90.5
REMARK 200 DATA REDUNDANCY IN SHELL : 6.20
REMARK 200 R MERGE FOR SHELL (I) : 0.18100
REMARK 200 R SYM FOR SHELL (I) : NULL
REMARK 200 FOR SHELL : 14.300
REMARK 200
REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH; MAD
REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MAD
REMARK 200 SOFTWARE USED: SOLVE/RESOLVE AND ARP/WARP
REMARK 200 STARTING MODEL: NULL
REMARK 200
REMARK 200 REMARK: NULL
REMARK 280
REMARK 280 CRYSTAL
REMARK 280 SOLVENT CONTENT, VS (%): 45.00
REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.24
REMARK 280
REMARK 280 CRYSTALLIZATION CONDITIONS: LITHIUM SULFATE, PEG400, DIOXANE,
REMARK 280 HEPES, PH 7.5, VAPOR DIFFUSION, HANGING DROP, TEMPERATURE 95K
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 89.52250
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 89.52250
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 27.82700
REMARK 290 SMTRY2 5 0.000000 1.000000 0.000000 29.10250
REMARK 290 SMTRY3 5 0.000000 0.000000 1.000000 0.00000
REMARK 290 SMTRY1 6 -1.000000 0.000000 0.000000 27.82700
REMARK 290 SMTRY2 6 0.000000 -1.000000 0.000000 29.10250
REMARK 290 SMTRY3 6 0.000000 0.000000 1.000000 89.52250
REMARK 290 SMTRY1 7 -1.000000 0.000000 0.000000 27.82700
REMARK 290 SMTRY2 7 0.000000 1.000000 0.000000 29.10250
REMARK 290 SMTRY3 7 0.000000 0.000000 -1.000000 89.52250
REMARK 290 SMTRY1 8 1.000000 0.000000 0.000000 27.82700
REMARK 290 SMTRY2 8 0.000000 -1.000000 0.000000 29.10250
REMARK 290 SMTRY3 8 0.000000 0.000000 -1.000000 0.00000
REMARK 290
REMARK 290 REMARK: NULL
REMARK 300
REMARK 300 BIOMOLECULE: 1
REMARK 300 THIS ENTRY CONTAINS THE CRYSTALLOGRAPHIC ASYMMETRIC UNIT
REMARK 300 WHICH CONSISTS OF 1 CHAIN(S). SEE REMARK 350 FOR
REMARK 300 INFORMATION ON GENERATING THE BIOLOGICAL MOLECULE(S).
REMARK 350
REMARK 350 GENERATING THE BIOMOLECULE
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 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 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 MET A 2
REMARK 465 LYS A 3
REMARK 465 ILE A 4
REMARK 465 VAL A 5
REMARK 525
REMARK 525 SOLVENT
REMARK 525 THE FOLLOWING SOLVENT MOLECULES LIE FARTHER THAN EXPECTED
REMARK 525 FROM THE PROTEIN OR NUCLEIC ACID MOLECULE AND MAY BE
REMARK 525 ASSOCIATED WITH A SYMMETRY RELATED MOLECULE (M=MODEL
REMARK 525 NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE
REMARK 525 NUMBER; I=INSERTION CODE):
REMARK 525
REMARK 525 M RES CSSEQI
REMARK 525 HOH 1119 DISTANCE = 6.11 ANGSTROMS
DBREF 1TQH A 1 247 SWS Q06174 EST_BACST 1 247
SEQRES 1 A 247 MET MET LYS ILE VAL PRO PRO LYS PRO PHE PHE PHE GLU
SEQRES 2 A 247 ALA GLY GLU ARG ALA VAL LEU LEU LEU HIS GLY PHE THR
SEQRES 3 A 247 GLY ASN SER ALA ASP VAL ARG MET LEU GLY ARG PHE LEU
SEQRES 4 A 247 GLU SER LYS GLY TYR THR CYS HIS ALA PRO ILE TYR LYS
SEQRES 5 A 247 GLY HIS GLY VAL PRO PRO GLU GLU LEU VAL HIS THR GLY
SEQRES 6 A 247 PRO ASP ASP TRP TRP GLN ASP VAL MET ASN GLY TYR GLU
SEQRES 7 A 247 PHE LEU LYS ASN LYS GLY TYR GLU LYS ILE ALA VAL ALA
SEQRES 8 A 247 GLY LEU SER LEU GLY GLY VAL PHE SER LEU LYS LEU GLY
SEQRES 9 A 247 TYR THR VAL PRO ILE GLU GLY ILE VAL THR MET CYS ALA
SEQRES 10 A 247 PRO MET TYR ILE LYS SER GLU GLU THR MET TYR GLU GLY
SEQRES 11 A 247 VAL LEU GLU TYR ALA ARG GLU TYR LYS LYS ARG GLU GLY
SEQRES 12 A 247 LYS SER GLU GLU GLN ILE GLU GLN GLU MET GLU LYS PHE
SEQRES 13 A 247 LYS GLN THR PRO MET LYS THR LEU LYS ALA LEU GLN GLU
SEQRES 14 A 247 LEU ILE ALA ASP VAL ARG ASP HIS LEU ASP LEU ILE TYR
SEQRES 15 A 247 ALA PRO THR PHE VAL VAL GLN ALA ARG HIS ASP GLU MET
SEQRES 16 A 247 ILE ASN PRO ASP SER ALA ASN ILE ILE TYR ASN GLU ILE
SEQRES 17 A 247 GLU SER PRO VAL LYS GLN ILE LYS TRP TYR GLU GLN SER
SEQRES 18 A 247 GLY HIS VAL ILE THR LEU ASP GLN GLU LYS ASP GLN LEU
SEQRES 19 A 247 HIS GLU ASP ILE TYR ALA PHE LEU GLU SER LEU ASP TRP
HET SO4 501 5
HET SO4 502 5
HET 4PA A 701 7
HETNAM SO4 SULFATE ION
HETNAM 4PA PROPYL ACETATE
FORMUL 2 SO4 2(O4 S1 2-)
FORMUL 4 4PA C5 H10 O2
FORMUL 5 HOH *153(H2 O1)
HELIX 1 1 SER A 29 LYS A 42 1 14
HELIX 2 2 PRO A 57 VAL A 62 1 6
HELIX 3 3 GLY A 65 LYS A 83 1 19
HELIX 4 4 SER A 94 TYR A 105 1 12
HELIX 5 5 SER A 123 GLY A 143 1 21
HELIX 6 6 SER A 145 LYS A 157 1 13
HELIX 7 7 THR A 163 HIS A 177 1 15
HELIX 8 8 LEU A 178 ILE A 181 5 4
HELIX 9 9 ASP A 199 ILE A 208 1 10
HELIX 10 10 VAL A 224 ASP A 228 5 5
HELIX 11 11 GLU A 230 LEU A 245 1 16
SHEET 1 A 7 PHE A 10 PHE A 12 0
SHEET 2 A 7 THR A 45 ALA A 48 -1 O CYS A 46 N PHE A 12
SHEET 3 A 7 ALA A 18 LEU A 22 1 N LEU A 21 O HIS A 47
SHEET 4 A 7 ILE A 88 LEU A 93 1 O ALA A 91 N LEU A 20
SHEET 5 A 7 ILE A 112 MET A 115 1 O MET A 115 N GLY A 92
SHEET 6 A 7 THR A 185 ALA A 190 1 O VAL A 188 N THR A 114
SHEET 7 A 7 LYS A 213 TYR A 218 1 O GLN A 214 N VAL A 187
LINK OG SER A 94 CAI 4PA A 701
CRYST1 55.654 58.205 179.045 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 0.017968 0.000000 0.000000 0.00000
SCALE2 0.000000 0.017181 0.000000 0.00000
SCALE3 0.000000 0.000000 0.005585 0.00000
MASTER 213 0 3 11 7 0 0 6 2168 1 18 19
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