LinkDB: 1E5H 1E5I
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HEADER OXIDOREDUCTASE 26-JUL-00 1E5H TITLE DELTA-R307A DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH SUCCINATE TITLE 2 AND CARBON DIOXIDE COMPND MOL_ID: 1; COMPND 2 MOLECULE: DEACETOXYCEPHALOSPORIN C SYNTHASE; COMPND 3 CHAIN: A; COMPND 4 SYNONYM: RING EXPANDING ENZYME, RING EXPANDASE; COMPND 5 ENGINEERED: YES; COMPND 6 MUTATION: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: STREPTOMYCES CLAVULIGERUS; SOURCE 3 ORGANISM_TAXID: 1901; SOURCE 4 GENE: CEFE; SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 6 EXPRESSION_SYSTEM_TAXID: 469008; SOURCE 7 EXPRESSION_SYSTEM_STRAIN: BL21(DE3); SOURCE 8 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID; SOURCE 9 EXPRESSION_SYSTEM_VECTOR: PET; SOURCE 10 EXPRESSION_SYSTEM_PLASMID: PET 24A; SOURCE 11 OTHER_DETAILS: RECOMBINANT E.COLI KEYWDS FERROUS OXYGENASE, CEPHALOSPORIN, 2-OXOGLUTARATE, C-TERMINUS KEYWDS 2 ANTIBIOTICS, OXIDOREDUCTASE, OXIDATIVE COUPLING CONTROL EXPDTA X-RAY DIFFRACTION AUTHOR H.J.LEE,M.D.LLOYD,K.HARLOS,I.J.CLIFTON,J.E.BALDWIN,C.J.SCHOFIELD REVDAT 4 13-DEC-23 1E5H 1 REMARK LINK REVDAT 3 24-FEB-09 1E5H 1 VERSN REVDAT 2 24-JUN-03 1E5H 1 REMARK HET HETNAM FORMUL REVDAT 2 2 1 LINK CONECT REVDAT 1 26-JUL-01 1E5H 0 JRNL AUTH H.J.LEE,M.D.LLOYD,K.HARLOS,I.J.CLIFTON,J.E.BALDWIN, JRNL AUTH 2 C.J.SCHOFIELD JRNL TITL KINETIC AND CRYSTALLOGRAPHIC STUDIES ON JRNL TITL 2 DEACETOXYCEPHALOSPORIN C SYNTHASE (DAOCS) JRNL REF J.MOL.BIOL. V. 308 937 2001 JRNL REFN ISSN 0022-2836 JRNL PMID 11352583 JRNL DOI 10.1006/JMBI.2001.4649 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH M.D.LLOYD,H.J.LEE,K.HARLOS,Z.-H.ZHANG,J.E.BALDWIN, REMARK 1 AUTH 2 C.J.SCHOFIELD,J.M.CHARNOCK,C.D.GARNER,T.HARA, REMARK 1 AUTH 3 A.C.TERRWISSCHA VAN SCHELTINGA,K.VALEGARD,J.A.C.VIKLUND, REMARK 1 AUTH 4 J.HAJDU,I.ANDERSSON,A.DANIELSSON,R.BHIKHABHAI REMARK 1 TITL STUDIES ON THE ACTIVE SITE OF DEACETOXYCEPHALOSPORIN C REMARK 1 TITL 2 SYNTHASE (DAOCS) REMARK 1 REF J.MOL.BIOL. V. 287 943 1999 REMARK 1 REFN ISSN 0022-2836 REMARK 1 PMID 10222202 REMARK 1 DOI 10.1006/JMBI.1999.2594 REMARK 2 REMARK 2 RESOLUTION. 1.96 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : CNS 0.9 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 : NULL REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 1.96 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 16.57 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.000 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 2006850.480 REMARK 3 DATA CUTOFF LOW (ABS(F)) : NULL REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 99.8 REMARK 3 NUMBER OF REFLECTIONS : 21428 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) : 0.220 REMARK 3 FREE R VALUE : 0.252 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.300 REMARK 3 FREE R VALUE TEST SET COUNT : 912 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) : 1.96 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.08 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 99.80 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 3454 REMARK 3 BIN R VALUE (WORKING SET) : 0.2630 REMARK 3 BIN FREE R VALUE : 0.3000 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 3.80 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 137 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.026 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 2126 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 12 REMARK 3 SOLVENT ATOMS : 119 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 21.60 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 33.60 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : -1.39000 REMARK 3 B22 (A**2) : -1.39000 REMARK 3 B33 (A**2) : 2.79000 REMARK 3 B12 (A**2) : 0.75000 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) : 0.25 REMARK 3 ESD FROM SIGMAA (A) : 0.17 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) : 0.31 REMARK 3 ESD FROM C-V SIGMAA (A) : 0.22 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES. REMARK 3 BOND LENGTHS (A) : 0.012 REMARK 3 BOND ANGLES (DEGREES) : 1.300 REMARK 3 DIHEDRAL ANGLES (DEGREES) : 22.70 REMARK 3 IMPROPER ANGLES (DEGREES) : 0.710 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) : 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 BULK SOLVENT MODELING. REMARK 3 METHOD USED : FLAT MODEL REMARK 3 KSOL : 0.39 REMARK 3 BSOL : 47.89 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 : SUCCINATE.PAR REMARK 3 PARAMETER FILE 5 : NULL REMARK 3 TOPOLOGY FILE 1 : PROTEIN.TOP REMARK 3 TOPOLOGY FILE 2 : ION.TOP REMARK 3 TOPOLOGY FILE 3 : WATER.TOP REMARK 3 TOPOLOGY FILE 4 : SUCCINATE.TOP REMARK 3 TOPOLOGY FILE 5 : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: DATA DETWINNED USING CCP4 PROGRAMME REMARK 3 DETWIN (A. LESLIE) REMARK 4 REMARK 4 1E5H COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE ON 26-JUL-00. REMARK 100 THE DEPOSITION ID IS D_1290005192. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 28-DEC-98 REMARK 200 TEMPERATURE (KELVIN) : 100.0 REMARK 200 PH : 7.00 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : N REMARK 200 RADIATION SOURCE : ROTATING ANODE REMARK 200 BEAMLINE : NULL REMARK 200 X-RAY GENERATOR MODEL : RIGAKU REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 1.5418 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : YALE MIRRORS REMARK 200 REMARK 200 DETECTOR TYPE : IMAGE PLATE REMARK 200 DETECTOR MANUFACTURER : MAR SCANNER 300 MM PLATE REMARK 200 INTENSITY-INTEGRATION SOFTWARE : DENZO REMARK 200 DATA SCALING SOFTWARE : SCALEPACK REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 21766 REMARK 200 RESOLUTION RANGE HIGH (A) : 1.960 REMARK 200 RESOLUTION RANGE LOW (A) : 30.000 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 99.8 REMARK 200 DATA REDUNDANCY : 5.920 REMARK 200 R MERGE (I) : 0.09000 REMARK 200 R SYM (I) : NULL REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : 25.9000 REMARK 200 REMARK 200 IN THE HIGHEST RESOLUTION SHELL. REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 1.95 REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 1.98 REMARK 200 COMPLETENESS FOR SHELL (%) : 99.6 REMARK 200 DATA REDUNDANCY IN SHELL : NULL REMARK 200 R MERGE FOR SHELL (I) : 0.28100 REMARK 200 R SYM FOR SHELL (I) : NULL REMARK 200 <I/SIGMA(I)> FOR SHELL : 6.290 REMARK 200 REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT REMARK 200 SOFTWARE USED: CNS RIGID BODY REFINEMENT REMARK 200 STARTING MODEL: 1RXF REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 48.16 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.39 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 100 MM HEPES-NAOH, PH 7.0, 0.5 MM REMARK 280 PB(OAC)2(OH2)2, 5 MM NA2SUCCINATE, 5 MM NAHCO3, GLYCEROL 5-10% REMARK 280 (W/V), 1.4-1.6 M AMMONIUM SULPHATE, PH 7.00 REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: H 3 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -Y,X-Y,Z REMARK 290 3555 -X+Y,-X,Z REMARK 290 4555 X+2/3,Y+1/3,Z+1/3 REMARK 290 5555 -Y+2/3,X-Y+1/3,Z+1/3 REMARK 290 6555 -X+Y+2/3,-X+1/3,Z+1/3 REMARK 290 7555 X+1/3,Y+2/3,Z+2/3 REMARK 290 8555 -Y+1/3,X-Y+2/3,Z+2/3 REMARK 290 9555 -X+Y+1/3,-X+2/3,Z+2/3 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 -0.500000 -0.866025 0.000000 0.00000 REMARK 290 SMTRY2 2 0.866025 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 3 -0.500000 0.866025 0.000000 0.00000 REMARK 290 SMTRY2 3 -0.866025 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 53.13000 REMARK 290 SMTRY2 4 0.000000 1.000000 0.000000 30.67462 REMARK 290 SMTRY3 4 0.000000 0.000000 1.000000 23.68667 REMARK 290 SMTRY1 5 -0.500000 -0.866025 0.000000 53.13000 REMARK 290 SMTRY2 5 0.866025 -0.500000 0.000000 30.67462 REMARK 290 SMTRY3 5 0.000000 0.000000 1.000000 23.68667 REMARK 290 SMTRY1 6 -0.500000 0.866025 0.000000 53.13000 REMARK 290 SMTRY2 6 -0.866025 -0.500000 0.000000 30.67462 REMARK 290 SMTRY3 6 0.000000 0.000000 1.000000 23.68667 REMARK 290 SMTRY1 7 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 7 0.000000 1.000000 0.000000 61.34924 REMARK 290 SMTRY3 7 0.000000 0.000000 1.000000 47.37333 REMARK 290 SMTRY1 8 -0.500000 -0.866025 0.000000 0.00000 REMARK 290 SMTRY2 8 0.866025 -0.500000 0.000000 61.34924 REMARK 290 SMTRY3 8 0.000000 0.000000 1.000000 47.37333 REMARK 290 SMTRY1 9 -0.500000 0.866025 0.000000 0.00000 REMARK 290 SMTRY2 9 -0.866025 -0.500000 0.000000 61.34924 REMARK 290 SMTRY3 9 0.000000 0.000000 1.000000 47.37333 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 300 REMARK: BIOLOGICALUNIT: ACTIVE AS A MONOMER, TRIMERIC REMARK 300 ASSEMBLYOBSERVED IN THE CRYSTAL. 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 SOFTWARE DETERMINED QUATERNARY STRUCTURE: TRIMERIC REMARK 350 SOFTWARE USED: PQS REMARK 350 TOTAL BURIED SURFACE AREA: 8600 ANGSTROM**2 REMARK 350 SURFACE AREA OF THE COMPLEX: 37920 ANGSTROM**2 REMARK 350 CHANGE IN SOLVENT FREE ENERGY: -29.9 KCAL/MOL 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 BIOMT1 2 -0.500000 -0.866025 0.000000 0.00000 REMARK 350 BIOMT2 2 0.866025 -0.500000 0.000000 0.00000 REMARK 350 BIOMT3 2 0.000000 0.000000 1.000000 0.00000 REMARK 350 BIOMT1 3 -0.500000 0.866025 0.000000 0.00000 REMARK 350 BIOMT2 3 -0.866025 -0.500000 0.000000 0.00000 REMARK 350 BIOMT3 3 0.000000 0.000000 1.000000 0.00000 REMARK 400 REMARK 400 COMPOUND REMARK 400 CHAIN A ENGINEERED MUTATION THR308ALA. THE C-TERMINAL RESIDUES REMARK 400 SER, LYS, ALA ARE DELETED. REMARK 400 REMARK 400 FUNCTION: DAOCS CATALYZES THE STEP FROM PENICILLIN N TO DEACETOXY- REMARK 400 CEPHALOSPORIN C. REMARK 400 COFACTOR: IRON AND ASCORBATE. REMARK 400 PATHWAY: BIOSYNTHESIS OF CEPHALOSPORIN ANTIBIOTICS. REMARK 400 SIMILARITY: BELONGS TO THE IRONULLSCORBATE-DEPENDENT FAMILY OF REMARK 400 OXIDOREDUCTASES. STRONG, TO CEFF. 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 LEU A 80 REMARK 465 GLU A 81 REMARK 465 SER A 82 REMARK 465 GLU A 83 REMARK 465 SER A 84 REMARK 465 THR A 85 REMARK 465 ALA A 86 REMARK 465 GLN A 87 REMARK 465 ILE A 88 REMARK 465 THR A 89 REMARK 465 ASN A 90 REMARK 465 GLN A 166 REMARK 465 VAL A 167 REMARK 465 PRO A 168 REMARK 465 GLU A 169 REMARK 465 HIS A 170 REMARK 465 ARG A 171 REMARK 465 SER A 172 REMARK 465 ALA A 173 REMARK 465 GLU A 174 REMARK 465 GLU A 175 REMARK 465 GLN A 176 REMARK 465 PRO A 177 REMARK 465 PRO A 196 REMARK 465 CYS A 197 REMARK 465 ALA A 198 REMARK 465 ASN A 199 REMARK 465 GLY A 200 REMARK 465 PRO A 248 REMARK 465 ARG A 249 REMARK 465 ARG A 250 REMARK 465 ASP A 251 REMARK 465 GLN A 252 REMARK 465 ILE A 253 REMARK 465 ALA A 254 REMARK 465 GLY A 255 REMARK 465 SER A 256 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 MET A 180 CG SD CE REMARK 470 ARG A 258 CG CD NE CZ NH1 NH2 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 SER A 93 -160.10 -123.34 REMARK 500 SER A 98 138.28 174.43 REMARK 500 CYS A 155 -139.28 -120.70 REMARK 500 SER A 286 64.99 -115.39 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 FE2 A 901 FE REMARK 620 N RES CSSEQI ATOM REMARK 620 1 HIS A 183 NE2 REMARK 620 2 ASP A 185 OD1 82.8 REMARK 620 3 ASP A 185 OD2 134.2 51.7 REMARK 620 4 HIS A 243 NE2 85.4 91.2 90.0 REMARK 620 5 SIN A 902 O1 84.7 161.1 140.4 102.0 REMARK 620 6 CO2 A 903 O2 86.7 97.0 102.1 167.9 68.0 REMARK 620 7 HOH A2072 O 164.6 106.5 57.4 106.2 83.0 80.0 REMARK 620 N 1 2 3 4 5 6 REMARK 800 REMARK 800 SITE REMARK 800 SITE_IDENTIFIER: AC1 REMARK 800 EVIDENCE_CODE: SOFTWARE REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE FE2 A 901 REMARK 800 REMARK 800 SITE_IDENTIFIER: AC2 REMARK 800 EVIDENCE_CODE: SOFTWARE REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SIN A 902 REMARK 800 REMARK 800 SITE_IDENTIFIER: AC3 REMARK 800 EVIDENCE_CODE: SOFTWARE REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE CO2 A 903 REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: 1RXF RELATED DB: PDB REMARK 900 DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH FE(II) REMARK 900 RELATED ID: 1RXG RELATED DB: PDB REMARK 900 DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH FE(II) AND 2- REMARK 900 OXOGLUTARATE REMARK 900 RELATED ID: 1DCS RELATED DB: PDB REMARK 900 DEACETOXYCEPHALOSPORIN C SYNTHASE FROM S. CLAVULIGERUS REMARK 900 RELATED ID: 1E5I RELATED DB: PDB REMARK 900 DELTA-R306 DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH IRON REMARK 900 AND 2-OXOGLUTARATE. REMARK 999 REMARK 999 SEQUENCE REMARK 999 ILE A 50, WRONGLY REPORTED IN DATABASE DBREF 1E5H A 1 308 UNP P18548 CEFE_STRCL 1 308 SEQADV 1E5H ILE A 50 UNP P18548 LEU 50 CONFLICT SEQADV 1E5H ALA A 308 UNP P18548 THR 308 ENGINEERED MUTATION SEQRES 1 A 308 MET ASP THR THR VAL PRO THR PHE SER LEU ALA GLU LEU SEQRES 2 A 308 GLN GLN GLY LEU HIS GLN ASP GLU PHE ARG ARG CYS LEU SEQRES 3 A 308 ARG ASP LYS GLY LEU PHE TYR LEU THR ASP CYS GLY LEU SEQRES 4 A 308 THR ASP THR GLU LEU LYS SER ALA LYS ASP ILE VAL ILE SEQRES 5 A 308 ASP PHE PHE GLU HIS GLY SER GLU ALA GLU LYS ARG ALA SEQRES 6 A 308 VAL THR SER PRO VAL PRO THR MET ARG ARG GLY PHE THR SEQRES 7 A 308 GLY LEU GLU SER GLU SER THR ALA GLN ILE THR ASN THR SEQRES 8 A 308 GLY SER TYR SER ASP TYR SER MET CYS TYR SER MET GLY SEQRES 9 A 308 THR ALA ASP ASN LEU PHE PRO SER GLY ASP PHE GLU ARG SEQRES 10 A 308 ILE TRP THR GLN TYR PHE ASP ARG GLN TYR THR ALA SER SEQRES 11 A 308 ARG ALA VAL ALA ARG GLU VAL LEU ARG ALA THR GLY THR SEQRES 12 A 308 GLU PRO ASP GLY GLY VAL GLU ALA PHE LEU ASP CYS GLU SEQRES 13 A 308 PRO LEU LEU ARG PHE ARG TYR PHE PRO GLN VAL PRO GLU SEQRES 14 A 308 HIS ARG SER ALA GLU GLU GLN PRO LEU ARG MET ALA PRO SEQRES 15 A 308 HIS TYR ASP LEU SER MET VAL THR LEU ILE GLN GLN THR SEQRES 16 A 308 PRO CYS ALA ASN GLY PHE VAL SER LEU GLN ALA GLU VAL SEQRES 17 A 308 GLY GLY ALA PHE THR ASP LEU PRO TYR ARG PRO ASP ALA SEQRES 18 A 308 VAL LEU VAL PHE CYS GLY ALA ILE ALA THR LEU VAL THR SEQRES 19 A 308 GLY GLY GLN VAL LYS ALA PRO ARG HIS HIS VAL ALA ALA SEQRES 20 A 308 PRO ARG ARG ASP GLN ILE ALA GLY SER SER ARG THR SER SEQRES 21 A 308 SER VAL PHE PHE LEU ARG PRO ASN ALA ASP PHE THR PHE SEQRES 22 A 308 SER VAL PRO LEU ALA ARG GLU CYS GLY PHE ASP VAL SER SEQRES 23 A 308 LEU ASP GLY GLU THR ALA THR PHE GLN ASP TRP ILE GLY SEQRES 24 A 308 GLY ASN TYR VAL ASN ILE ARG ARG ALA HET FE2 A 901 1 HET SIN A 902 8 HET CO2 A 903 3 HETNAM FE2 FE (II) ION HETNAM SIN SUCCINIC ACID HETNAM CO2 CARBON DIOXIDE FORMUL 2 FE2 FE 2+ FORMUL 3 SIN C4 H6 O4 FORMUL 4 CO2 C O2 FORMUL 5 HOH *119(H2 O) HELIX 1 1 LEU A 10 GLN A 15 1 6 HELIX 2 2 HIS A 18 LYS A 29 1 12 HELIX 3 3 THR A 40 GLY A 58 1 19 HELIX 4 4 SER A 59 ALA A 65 1 7 HELIX 5 5 SER A 112 THR A 141 1 30 HELIX 6 6 GLY A 148 ASP A 154 1 7 HELIX 7 7 GLY A 227 THR A 234 1 8 HELIX 8 8 VAL A 275 CYS A 281 1 7 HELIX 9 9 PHE A 294 GLY A 299 1 6 SHEET 1 A 8 THR A 7 SER A 9 0 SHEET 2 A 8 LEU A 31 THR A 35 1 N TYR A 33 O PHE A 8 SHEET 3 A 8 VAL A 222 CYS A 226 -1 N VAL A 224 O PHE A 32 SHEET 4 A 8 VAL A 189 GLN A 194 -1 N ILE A 192 O LEU A 223 SHEET 5 A 8 THR A 259 LEU A 265 -1 N LEU A 265 O VAL A 189 SHEET 6 A 8 LEU A 158 PHE A 164 -1 N ARG A 162 O SER A 260 SHEET 7 A 8 SER A 98 MET A 103 -1 N MET A 103 O LEU A 159 SHEET 8 A 8 GLY A 76 THR A 78 -1 N THR A 78 O CYS A 100 SHEET 1 B 2 HIS A 183 ASP A 185 0 SHEET 2 B 2 ILE A 305 ARG A 307 -1 N ARG A 306 O TYR A 184 SHEET 1 C 3 HIS A 243 VAL A 245 0 SHEET 2 C 3 LEU A 204 GLU A 207 -1 N GLN A 205 O HIS A 244 SHEET 3 C 3 PHE A 212 ASP A 214 -1 N THR A 213 O ALA A 206 SHEET 1 D 2 THR A 272 SER A 274 0 SHEET 2 D 2 THR A 291 THR A 293 -1 N ALA A 292 O PHE A 273 LINK NE2 HIS A 183 FE FE2 A 901 1555 1555 2.31 LINK OD1 ASP A 185 FE FE2 A 901 1555 1555 2.14 LINK OD2 ASP A 185 FE FE2 A 901 1555 1555 2.69 LINK NE2 HIS A 243 FE FE2 A 901 1555 1555 2.30 LINK FE FE2 A 901 O1 SIN A 902 1555 1555 2.26 LINK FE FE2 A 901 O2 CO2 A 903 1555 1555 2.16 LINK FE FE2 A 901 O HOH A2072 1555 1555 2.14 SITE 1 AC1 6 HIS A 183 ASP A 185 HIS A 243 SIN A 902 SITE 2 AC1 6 CO2 A 903 HOH A2072 SITE 1 AC2 7 ARG A 162 HIS A 183 ILE A 192 HIS A 243 SITE 2 AC2 7 FE2 A 901 CO2 A 903 HOH A2072 SITE 1 AC3 8 ARG A 160 HIS A 183 ASP A 185 PHE A 264 SITE 2 AC3 8 ILE A 305 FE2 A 901 SIN A 902 HOH A2072 CRYST1 106.260 106.260 71.060 90.00 90.00 120.00 H 3 9 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 0.009411 0.005433 0.000000 0.00000 SCALE2 0.000000 0.010867 0.000000 0.00000 SCALE3 0.000000 0.000000 0.014073 0.00000 (ATOM LINES ARE NOT SHOWN.) END
HEADER OXIDOREDUCTASE 26-JUL-00 1E5I TITLE DELTA-R306 DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH IRON AND TITLE 2 2-OXOGLUTARATE. COMPND MOL_ID: 1; COMPND 2 MOLECULE: DEACETOXYCEPHALOSPORIN C SYNTHASE; COMPND 3 CHAIN: A; COMPND 4 FRAGMENT: RESIDUES 1-306; COMPND 5 SYNONYM: RING EXPANDING ENZYME, RING EXPANDASE, DAOCS, EXPANDASE; COMPND 6 EC: 1.14.20.1; COMPND 7 ENGINEERED: YES SOURCE MOL_ID: 1; SOURCE 2 ORGANISM_SCIENTIFIC: STREPTOMYCES CLAVULIGERUS; SOURCE 3 ORGANISM_TAXID: 1901; SOURCE 4 GENE: CEFE; SOURCE 5 EXPRESSION_SYSTEM: ESCHERICHIA COLI; SOURCE 6 EXPRESSION_SYSTEM_TAXID: 469008; SOURCE 7 EXPRESSION_SYSTEM_STRAIN: BL21(DE3); SOURCE 8 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID; SOURCE 9 EXPRESSION_SYSTEM_VECTOR: PET; SOURCE 10 EXPRESSION_SYSTEM_PLASMID: PET 24A; SOURCE 11 OTHER_DETAILS: RECOMBINANT E.COLI KEYWDS FERROUS OXYGENASE, CEPHALOSPORIN, 2-OXOGLUTARATE, C-TERMINUS KEYWDS 2 ANTIBIOTICS, OXIDOREDUCTASE, OXIDATIVE COUPLING CONTROL EXPDTA X-RAY DIFFRACTION AUTHOR H.J.LEE,M.D.LLOYD,K.HARLOS,I.J.CLIFTON,J.E.BALDWIN,C.J.SCHOFIELD REVDAT 6 13-DEC-23 1E5I 1 REMARK LINK REVDAT 5 24-AUG-11 1E5I 1 COMPND SOURCE REMARK HETSYN REVDAT 5 2 1 ATOM VERSN REVDAT 4 08-DEC-09 1E5I 1 VERSN REVDAT 3 24-FEB-09 1E5I 1 VERSN REVDAT 2 23-NOV-01 1E5I 1 JRNL REVDAT 1 26-JUL-01 1E5I 0 JRNL AUTH H.J.LEE,M.D.LLOYD,K.HARLOS,I.J.CLIFTON,J.E.BALDWIN, JRNL AUTH 2 C.J.SCHOFIELD JRNL TITL KINETIC AND CRYSTALLOGRAPHIC STUDIES ON JRNL TITL 2 DEACETOXYCEPHALOSPORIN C SYNTHASE (DAOCS) JRNL REF J.MOL.BIOL. V. 308 937 2001 JRNL REFN ISSN 0022-2836 JRNL PMID 11352583 JRNL DOI 10.1006/JMBI.2001.4649 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH M.D.LLOYD,H.J.LEE,K.HARLOS,Z.-H.ZHANG,J.E.BALDWIN, REMARK 1 AUTH 2 C.J.SCHOFIELD,J.M.CHARNOCK,C.D.GARNER,T.HARA, REMARK 1 AUTH 3 A.C.TERRWISSCHA VAN SCHELTINGA,K.VALEGARD,J.A.C.VIKLUND, REMARK 1 AUTH 4 J.HAJDU,I.ANDERSSON,A.DANIELSSON,R.BHIKHABHAI REMARK 1 TITL STUDIES ON THE ACTIVE SITE OF DEACETOXYCEPHALOSPORIN C REMARK 1 TITL 2 SYNTHASE (DAOCS) REMARK 1 REF J.MOL.BIOL. V. 287 943 1999 REMARK 1 REFN ISSN 0022-2836 REMARK 1 PMID 10222202 REMARK 1 DOI 10.1006/JMBI.1999.2594 REMARK 2 REMARK 2 RESOLUTION. 2.10 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM : CNS 0.9 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 : NULL REMARK 3 REMARK 3 DATA USED IN REFINEMENT. REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.10 REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 19.99 REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.000 REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 2222669.750 REMARK 3 DATA CUTOFF LOW (ABS(F)) : NULL REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 98.8 REMARK 3 NUMBER OF REFLECTIONS : 17139 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) : 0.227 REMARK 3 FREE R VALUE : 0.242 REMARK 3 FREE R VALUE TEST SET SIZE (%) : 4.100 REMARK 3 FREE R VALUE TEST SET COUNT : 709 REMARK 3 ESTIMATED ERROR OF FREE R VALUE : 0.009 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) : 2.10 REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.23 REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 100.0 REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 2755 REMARK 3 BIN R VALUE (WORKING SET) : 0.2590 REMARK 3 BIN FREE R VALUE : 0.2780 REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 4.70 REMARK 3 BIN FREE R VALUE TEST SET COUNT : 137 REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.024 REMARK 3 REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT. REMARK 3 PROTEIN ATOMS : 2077 REMARK 3 NUCLEIC ACID ATOMS : 0 REMARK 3 HETEROGEN ATOMS : 11 REMARK 3 SOLVENT ATOMS : 123 REMARK 3 REMARK 3 B VALUES. REMARK 3 FROM WILSON PLOT (A**2) : 21.60 REMARK 3 MEAN B VALUE (OVERALL, A**2) : 32.80 REMARK 3 OVERALL ANISOTROPIC B VALUE. REMARK 3 B11 (A**2) : -1.10000 REMARK 3 B22 (A**2) : -1.10000 REMARK 3 B33 (A**2) : 2.20000 REMARK 3 B12 (A**2) : 1.22000 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) : 0.26 REMARK 3 ESD FROM SIGMAA (A) : 0.20 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) : 0.29 REMARK 3 ESD FROM C-V SIGMAA (A) : 0.21 REMARK 3 REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES. REMARK 3 BOND LENGTHS (A) : 0.012 REMARK 3 BOND ANGLES (DEGREES) : 1.300 REMARK 3 DIHEDRAL ANGLES (DEGREES) : 23.10 REMARK 3 IMPROPER ANGLES (DEGREES) : 0.720 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) : 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 BULK SOLVENT MODELING. REMARK 3 METHOD USED : FLAT MODEL REMARK 3 KSOL : 0.39 REMARK 3 BSOL : 53.83 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 : 2-OXOGLUTARATE.PAR REMARK 3 PARAMETER FILE 5 : NULL REMARK 3 TOPOLOGY FILE 1 : PROTEIN.TOP REMARK 3 TOPOLOGY FILE 2 : ION.TOP REMARK 3 TOPOLOGY FILE 3 : WATER.TOP REMARK 3 TOPOLOGY FILE 4 : 2-OXOGLUTARATE.TOP REMARK 3 TOPOLOGY FILE 5 : NULL REMARK 3 REMARK 3 OTHER REFINEMENT REMARKS: DATA DETWINNED USING CCP4 PROGRAMME REMARK 3 DETWIN (A. LESLIE) REMARK 4 REMARK 4 1E5I COMPLIES WITH FORMAT V. 3.30, 13-JUL-11 REMARK 100 REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY PDBE ON 26-JUL-00. REMARK 100 THE DEPOSITION ID IS D_1290005193. REMARK 200 REMARK 200 EXPERIMENTAL DETAILS REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION REMARK 200 DATE OF DATA COLLECTION : 26-MAR-99 REMARK 200 TEMPERATURE (KELVIN) : 100.0 REMARK 200 PH : 7.00 REMARK 200 NUMBER OF CRYSTALS USED : 1 REMARK 200 REMARK 200 SYNCHROTRON (Y/N) : N REMARK 200 RADIATION SOURCE : ROTATING ANODE REMARK 200 BEAMLINE : NULL REMARK 200 X-RAY GENERATOR MODEL : RIGAKU REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M REMARK 200 WAVELENGTH OR RANGE (A) : 1.5418 REMARK 200 MONOCHROMATOR : NULL REMARK 200 OPTICS : YALE MIRRORS REMARK 200 REMARK 200 DETECTOR TYPE : IMAGE PLATE REMARK 200 DETECTOR MANUFACTURER : MAR SCANNER 345 MM PLATE REMARK 200 INTENSITY-INTEGRATION SOFTWARE : MOSFLM REMARK 200 DATA SCALING SOFTWARE : SCALA REMARK 200 REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 17034 REMARK 200 RESOLUTION RANGE HIGH (A) : 2.100 REMARK 200 RESOLUTION RANGE LOW (A) : 30.000 REMARK 200 REJECTION CRITERIA (SIGMA(I)) : NULL REMARK 200 REMARK 200 OVERALL. REMARK 200 COMPLETENESS FOR RANGE (%) : 99.9 REMARK 200 DATA REDUNDANCY : 3.200 REMARK 200 R MERGE (I) : 0.10800 REMARK 200 R SYM (I) : 0.11800 REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : 3.9800 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) : 2.11 REMARK 200 COMPLETENESS FOR SHELL (%) : 100.0 REMARK 200 DATA REDUNDANCY IN SHELL : 3.20 REMARK 200 R MERGE FOR SHELL (I) : 0.42300 REMARK 200 R SYM FOR SHELL (I) : 0.56100 REMARK 200 <I/SIGMA(I)> FOR SHELL : 6.290 REMARK 200 REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT REMARK 200 SOFTWARE USED: CNS RIGID BODY REFINEMENT REMARK 200 STARTING MODEL: PDB ENTRY 1RXF REMARK 200 REMARK 200 REMARK: NULL REMARK 280 REMARK 280 CRYSTAL REMARK 280 SOLVENT CONTENT, VS (%): 48.78 REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 2.42 REMARK 280 REMARK 280 CRYSTALLIZATION CONDITIONS: 100 MM HEPES-NAOH, PH 7.0, 0.5 MM REMARK 280 PB(OAC)2(OH2)2, 5 MM 2-OXOGLUTARATE, GLYCEROL 5-10% (W/V), 1.4- REMARK 280 1.65 M AMMONIUM SULPHATE, PH 7.00 REMARK 290 REMARK 290 CRYSTALLOGRAPHIC SYMMETRY REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: H 3 REMARK 290 REMARK 290 SYMOP SYMMETRY REMARK 290 NNNMMM OPERATOR REMARK 290 1555 X,Y,Z REMARK 290 2555 -Y,X-Y,Z REMARK 290 3555 -X+Y,-X,Z REMARK 290 4555 X+2/3,Y+1/3,Z+1/3 REMARK 290 5555 -Y+2/3,X-Y+1/3,Z+1/3 REMARK 290 6555 -X+Y+2/3,-X+1/3,Z+1/3 REMARK 290 7555 X+1/3,Y+2/3,Z+2/3 REMARK 290 8555 -Y+1/3,X-Y+2/3,Z+2/3 REMARK 290 9555 -X+Y+1/3,-X+2/3,Z+2/3 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 -0.500000 -0.866025 0.000000 0.00000 REMARK 290 SMTRY2 2 0.866025 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 3 -0.500000 0.866025 0.000000 0.00000 REMARK 290 SMTRY2 3 -0.866025 -0.500000 0.000000 0.00000 REMARK 290 SMTRY3 3 0.000000 0.000000 1.000000 0.00000 REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 52.89350 REMARK 290 SMTRY2 4 0.000000 1.000000 0.000000 30.53808 REMARK 290 SMTRY3 4 0.000000 0.000000 1.000000 23.74067 REMARK 290 SMTRY1 5 -0.500000 -0.866025 0.000000 52.89350 REMARK 290 SMTRY2 5 0.866025 -0.500000 0.000000 30.53808 REMARK 290 SMTRY3 5 0.000000 0.000000 1.000000 23.74067 REMARK 290 SMTRY1 6 -0.500000 0.866025 0.000000 52.89350 REMARK 290 SMTRY2 6 -0.866025 -0.500000 0.000000 30.53808 REMARK 290 SMTRY3 6 0.000000 0.000000 1.000000 23.74067 REMARK 290 SMTRY1 7 1.000000 0.000000 0.000000 0.00000 REMARK 290 SMTRY2 7 0.000000 1.000000 0.000000 61.07615 REMARK 290 SMTRY3 7 0.000000 0.000000 1.000000 47.48133 REMARK 290 SMTRY1 8 -0.500000 -0.866025 0.000000 0.00000 REMARK 290 SMTRY2 8 0.866025 -0.500000 0.000000 61.07615 REMARK 290 SMTRY3 8 0.000000 0.000000 1.000000 47.48133 REMARK 290 SMTRY1 9 -0.500000 0.866025 0.000000 0.00000 REMARK 290 SMTRY2 9 -0.866025 -0.500000 0.000000 61.07615 REMARK 290 SMTRY3 9 0.000000 0.000000 1.000000 47.48133 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 SOFTWARE DETERMINED QUATERNARY STRUCTURE: TRIMERIC REMARK 350 SOFTWARE USED: PISA REMARK 350 TOTAL BURIED SURFACE AREA: 7330 ANGSTROM**2 REMARK 350 SURFACE AREA OF THE COMPLEX: 29400 ANGSTROM**2 REMARK 350 CHANGE IN SOLVENT FREE ENERGY: -57.9 KCAL/MOL 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 BIOMT1 2 -0.500000 -0.866025 0.000000 0.00000 REMARK 350 BIOMT2 2 0.866025 -0.500000 0.000000 0.00000 REMARK 350 BIOMT3 2 0.000000 0.000000 1.000000 0.00000 REMARK 350 BIOMT1 3 -0.500000 0.866025 0.000000 0.00000 REMARK 350 BIOMT2 3 -0.866025 -0.500000 0.000000 0.00000 REMARK 350 BIOMT3 3 0.000000 0.000000 1.000000 0.00000 REMARK 350 REMARK 350 BIOMOLECULE: 2 REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: MONOMERIC 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 400 REMARK 400 COMPOUND REMARK 400 THE C-TERMINAL RESIDUES THR, SER, LYS, ALA ARE DELETED. REMARK 400 REMARK 400 FUNCTION: DAOCS CATALYZES THE STEP FROM PENICILLIN N TO DEACETOXY- REMARK 400 CEPHALOSPORIN C. REMARK 400 COFACTOR: IRON AND ASCORBATE. REMARK 400 PATHWAY: BIOSYNTHESIS OF CEPHALOSPORIN ANTIBIOTICS. REMARK 400 SIMILARITY: BELONGS TO THE IRONULLSCORBATE-DEPENDENT FAMILY OF REMARK 400 OXIDOREDUCTASES. STRONG, TO CEFF. 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 GLU A 81 REMARK 465 SER A 82 REMARK 465 GLU A 83 REMARK 465 SER A 84 REMARK 465 THR A 85 REMARK 465 ALA A 86 REMARK 465 GLN A 87 REMARK 465 ILE A 88 REMARK 465 THR A 89 REMARK 465 ASN A 90 REMARK 465 THR A 91 REMARK 465 GLY A 92 REMARK 465 SER A 93 REMARK 465 TYR A 94 REMARK 465 SER A 95 REMARK 465 ASP A 96 REMARK 465 TYR A 97 REMARK 465 VAL A 167 REMARK 465 PRO A 168 REMARK 465 GLU A 169 REMARK 465 HIS A 170 REMARK 465 ARG A 171 REMARK 465 SER A 172 REMARK 465 ALA A 173 REMARK 465 GLU A 174 REMARK 465 GLU A 175 REMARK 465 GLN A 176 REMARK 465 PRO A 177 REMARK 465 ASN A 199 REMARK 465 GLY A 200 REMARK 465 ARG A 249 REMARK 465 ARG A 250 REMARK 465 ASP A 251 REMARK 465 GLN A 252 REMARK 465 ILE A 253 REMARK 465 ALA A 254 REMARK 465 GLY A 255 REMARK 465 SER A 256 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 LEU A 80 CA C O CB CG CD1 CD2 REMARK 470 GLN A 166 CA C O CB CG CD OE1 REMARK 470 GLN A 166 NE2 REMARK 470 ALA A 198 CA C O CB REMARK 470 PRO A 248 CA C O CB CG CD 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 THR A 3 20.33 -78.10 REMARK 500 CYS A 155 -142.30 -118.16 REMARK 500 HIS A 183 167.00 176.35 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 FE2 A 901 FE REMARK 620 N RES CSSEQI ATOM REMARK 620 1 HIS A 183 NE2 REMARK 620 2 ASP A 185 OD1 79.6 REMARK 620 3 HIS A 243 NE2 91.2 90.2 REMARK 620 4 AKG A 902 O2 88.6 102.9 166.6 REMARK 620 5 AKG A 902 O5 100.1 171.0 98.7 68.1 REMARK 620 6 HOH A2074 O 174.1 105.6 85.9 93.0 75.3 REMARK 620 N 1 2 3 4 5 REMARK 800 REMARK 800 SITE REMARK 800 SITE_IDENTIFIER: AC1 REMARK 800 EVIDENCE_CODE: SOFTWARE REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE FE2 A 901 REMARK 800 REMARK 800 SITE_IDENTIFIER: AC2 REMARK 800 EVIDENCE_CODE: SOFTWARE REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE AKG A 902 REMARK 900 REMARK 900 RELATED ENTRIES REMARK 900 RELATED ID: 1RXG RELATED DB: PDB REMARK 900 DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH FE(II) AND 2- REMARK 900 OXOGLUTARATE REMARK 900 RELATED ID: 1RXF RELATED DB: PDB REMARK 900 DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH FE(II) REMARK 900 RELATED ID: 1DCS RELATED DB: PDB REMARK 900 DEACETOXYCEPHALOSPORIN C SYNTHASE FROM S. CLAVULIGERUS REMARK 900 RELATED ID: 1E5H RELATED DB: PDB REMARK 900 DELTA-R307A DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH REMARK 900 SUCCINATE AND CARBON DIOXIDE REMARK 999 REMARK 999 SEQUENCE REMARK 999 ILE A 50, WRONGLY REPORTED IN DATABASE DBREF 1E5I A 1 306 UNP P18548 CEFE_STRCL 1 306 SEQADV 1E5I ILE A 50 UNP P18548 LEU 50 SEE REMARK 999 SEQRES 1 A 306 MET ASP THR THR VAL PRO THR PHE SER LEU ALA GLU LEU SEQRES 2 A 306 GLN GLN GLY LEU HIS GLN ASP GLU PHE ARG ARG CYS LEU SEQRES 3 A 306 ARG ASP LYS GLY LEU PHE TYR LEU THR ASP CYS GLY LEU SEQRES 4 A 306 THR ASP THR GLU LEU LYS SER ALA LYS ASP ILE VAL ILE SEQRES 5 A 306 ASP PHE PHE GLU HIS GLY SER GLU ALA GLU LYS ARG ALA SEQRES 6 A 306 VAL THR SER PRO VAL PRO THR MET ARG ARG GLY PHE THR SEQRES 7 A 306 GLY LEU GLU SER GLU SER THR ALA GLN ILE THR ASN THR SEQRES 8 A 306 GLY SER TYR SER ASP TYR SER MET CYS TYR SER MET GLY SEQRES 9 A 306 THR ALA ASP ASN LEU PHE PRO SER GLY ASP PHE GLU ARG SEQRES 10 A 306 ILE TRP THR GLN TYR PHE ASP ARG GLN TYR THR ALA SER SEQRES 11 A 306 ARG ALA VAL ALA ARG GLU VAL LEU ARG ALA THR GLY THR SEQRES 12 A 306 GLU PRO ASP GLY GLY VAL GLU ALA PHE LEU ASP CYS GLU SEQRES 13 A 306 PRO LEU LEU ARG PHE ARG TYR PHE PRO GLN VAL PRO GLU SEQRES 14 A 306 HIS ARG SER ALA GLU GLU GLN PRO LEU ARG MET ALA PRO SEQRES 15 A 306 HIS TYR ASP LEU SER MET VAL THR LEU ILE GLN GLN THR SEQRES 16 A 306 PRO CYS ALA ASN GLY PHE VAL SER LEU GLN ALA GLU VAL SEQRES 17 A 306 GLY GLY ALA PHE THR ASP LEU PRO TYR ARG PRO ASP ALA SEQRES 18 A 306 VAL LEU VAL PHE CYS GLY ALA ILE ALA THR LEU VAL THR SEQRES 19 A 306 GLY GLY GLN VAL LYS ALA PRO ARG HIS HIS VAL ALA ALA SEQRES 20 A 306 PRO ARG ARG ASP GLN ILE ALA GLY SER SER ARG THR SER SEQRES 21 A 306 SER VAL PHE PHE LEU ARG PRO ASN ALA ASP PHE THR PHE SEQRES 22 A 306 SER VAL PRO LEU ALA ARG GLU CYS GLY PHE ASP VAL SER SEQRES 23 A 306 LEU ASP GLY GLU THR ALA THR PHE GLN ASP TRP ILE GLY SEQRES 24 A 306 GLY ASN TYR VAL ASN ILE ARG HET FE2 A 901 1 HET AKG A 902 10 HETNAM FE2 FE (II) ION HETNAM AKG 2-OXOGLUTARIC ACID FORMUL 2 FE2 FE 2+ FORMUL 3 AKG C5 H6 O5 FORMUL 4 HOH *123(H2 O) HELIX 1 1 LEU A 10 GLN A 15 1 6 HELIX 2 2 HIS A 18 LYS A 29 1 12 HELIX 3 3 THR A 40 GLY A 58 1 19 HELIX 4 4 SER A 59 VAL A 66 1 8 HELIX 5 5 SER A 112 THR A 141 1 30 HELIX 6 6 GLY A 148 ASP A 154 1 7 HELIX 7 7 GLY A 227 THR A 234 1 8 HELIX 8 8 VAL A 275 CYS A 281 1 7 HELIX 9 9 PHE A 294 GLY A 299 1 6 SHEET 1 A 8 THR A 7 SER A 9 0 SHEET 2 A 8 LEU A 31 THR A 35 1 N TYR A 33 O PHE A 8 SHEET 3 A 8 VAL A 222 CYS A 226 -1 N VAL A 224 O PHE A 32 SHEET 4 A 8 VAL A 189 THR A 195 -1 N ILE A 192 O LEU A 223 SHEET 5 A 8 ARG A 258 LEU A 265 -1 N LEU A 265 O VAL A 189 SHEET 6 A 8 LEU A 158 PHE A 164 -1 N PHE A 164 O ARG A 258 SHEET 7 A 8 MET A 99 MET A 103 -1 N MET A 103 O LEU A 159 SHEET 8 A 8 GLY A 76 THR A 78 -1 N THR A 78 O CYS A 100 SHEET 1 B 3 HIS A 243 VAL A 245 0 SHEET 2 B 3 LEU A 204 VAL A 208 -1 N GLN A 205 O HIS A 244 SHEET 3 B 3 ALA A 211 ASP A 214 -1 N THR A 213 O ALA A 206 SHEET 1 C 2 THR A 272 SER A 274 0 SHEET 2 C 2 THR A 291 THR A 293 -1 N ALA A 292 O PHE A 273 LINK NE2 HIS A 183 FE FE2 A 901 1555 1555 2.25 LINK OD1 ASP A 185 FE FE2 A 901 1555 1555 2.26 LINK NE2 HIS A 243 FE FE2 A 901 1555 1555 2.20 LINK FE FE2 A 901 O2 AKG A 902 1555 1555 2.15 LINK FE FE2 A 901 O5 AKG A 902 1555 1555 2.28 LINK FE FE2 A 901 O HOH A2074 1555 1555 2.32 SITE 1 AC1 5 HIS A 183 ASP A 185 HIS A 243 AKG A 902 SITE 2 AC1 5 HOH A2074 SITE 1 AC2 11 ARG A 162 PHE A 164 HIS A 183 ASP A 185 SITE 2 AC2 11 HIS A 243 ARG A 258 SER A 260 PHE A 264 SITE 3 AC2 11 ILE A 305 FE2 A 901 HOH A2074 CRYST1 105.787 105.787 71.222 90.00 90.00 120.00 H 3 9 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 0.009453 0.005458 0.000000 0.00000 SCALE2 0.000000 0.010915 0.000000 0.00000 SCALE3 0.000000 0.000000 0.014041 0.00000 (ATOM LINES ARE NOT SHOWN.) END