HEADER OXIDOREDUCTASE(H2O2(A)) 07-JUN-93 7CCP COMPND CYTOCHROME C PEROXIDASE (E.C.1.11.1.5) MUTANT WITH MET ILE COMPND 2 ADDED AT N-TERMINUS AND ARG 48 REPLACED BY LEU (MI,R48L) SOURCE YEAST (SACCHAROMYCES CEREVISIAE) RECOMBINANT FORM EXPRESSED SOURCE 2 IN (ESCHERICHIA COLI) AUTHOR J.WANG,M.A.MILLER,J.KRAUT REVDAT 1 31-OCT-93 7CCP 0 JRNL AUTH L.B.VITELLO,J.E.ERMAN,M.A.MILLER,J.WANG,J.KRAUT JRNL TITL EFFECT OF ARGININE-48 REPLACEMENT ON THE REACTION JRNL TITL 2 BETWEEN CYTOCHROME C PEROXIDASE AND HYDROGEN JRNL TITL 3 PEROXIDE JRNL REF BIOCHEMISTRY V. 32 9807 1993 JRNL REFN ASTM BICHAW US ISSN 0006-2960 033 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH J.WANG,J.M.MAURO,S.L.EDWARDS,S.J.OATLEY,L.A.FISHEL, REMARK 1 AUTH 2 V.A.ASHFORD,N.-H.XUONG,J.KRAUT REMARK 1 TITL X-RAY STRUCTURES OF RECOMBINANT YEAST CYTOCHROME REMARK 1 TITL 2 C PEROXIDASE AND THREE HEME CLEFT MUTANTS REMARK 1 TITL 3 PREPARED BY SITE-DIRECTED MUTAGENESIS REMARK 1 REF BIOCHEMISTRY V. 29 7160 1990 REMARK 1 REFN ASTM BICHAW US ISSN 0006-2960 033 REMARK 1 REFERENCE 2 REMARK 1 AUTH B.C.FINZEL,T.L.POULOS,J.KRAUT REMARK 1 TITL CRYSTAL STRUCTURE OF YEAST CYTOCHROME C REMARK 1 TITL 2 PEROXIDASE REFINED AT 1.7 ANGSTROMS RESOLUTION REMARK 1 REF J.BIOL.CHEM. V. 259 13027 1984 REMARK 1 REFN ASTM JBCHA3 US ISSN 0021-9258 071 REMARK 2 REMARK 2 RESOLUTION. 2.2 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM PROLSQ REMARK 3 AUTHORS KONNERT,HENDRICKSON REMARK 3 R VALUE 0.159 REMARK 3 REMARK 3 NUMBER OF REFLECTIONS 15996 REMARK 3 REMARK 3 NUMBER OF PROTEIN ATOMS 2365 REMARK 3 NUMBER OF SOLVENT ATOMS 199 REMARK 3 REMARK 3 DISTANCE RESTRAINTS (ANGSTROMS) REMARK 3 BOND DISTANCE 0.012 REMARK 3 ANGLE DISTANCE 0.028 REMARK 3 PLANAR 1-4 DISTANCE 0.034 REMARK 3 PLANE RESTRAINT (ANGSTROMS) 0.012 REMARK 3 CHIRAL-CENTER RESTRAINT (ANGSTROMS**3) 0.15 REMARK 3 CONFORMATIONAL TORSION ANGLE RESTRAINT (DEGREES) REMARK 3 PLANAR 2.7 REMARK 3 STAGGERED 16.4 REMARK 4 REMARK 4 THIS CYTOCHROME C PEROXIDASE DIFFERS FROM A PREVIOUSLY REMARK 4 DEPOSITED STRUCTURE (PROTEIN DATA BANK ENTRY 2CYP) BY REMARK 4 TWO STRAIN-RELATED SEQUENCE DIFFERENCES - THR 53 TO ILE REMARK 4 AND ASP 152 TO GLY, AND THE ADDITION OF MET-ILE AT THE REMARK 4 N-TERMINUS, HENCE CCP(MI). THE OVERALL STRUCTURE IS THE REMARK 4 SAME AS THE PREVIOUSLY DEPOSITED ONE BUT IN A DIFFERENT REMARK 4 SPACE GROUP. REMARK 5 REMARK 5 THE SEQUENCE DIFFERS FROM THAT REPORTED FOR 2CYP AT RESIDUE REMARK 5 272. IN THE PRESENT ENTRY, THIS RESIDUE IS REPORTED AS REMARK 5 ASN. THIS CORRECTS AN ERROR INTRODUCED IN 2CYP, WHERE THE REMARK 5 RESIDUE IS INCORRECTLY REPORTED TO BE ASP. REMARK 6 REMARK 6 RESIDUES ARE NUMBERED TO BE CONSISTENT WITH THE SEQUENCE OF REMARK 6 THE NATIVE (2CYP) STRUCTURE. THUS THE FIRST TWO RESIDUES REMARK 6 HAVE RESIDUE NUMBERS -1 AND 0, RESPECTIVELY. REMARK 7 REMARK 7 COORDINATES FOR RESIDUES -1, 0, AND 1 - 3 ARE NOT INCLUDED REMARK 7 IN THIS ENTRY BECAUSE THESE RESIDUES COULD NOT BE RESOLVED REMARK 7 IN THE FINAL ELECTRON DENSITY MAPS. REMARK 8 REMARK 8 THE NATIVE STRUCTURE IS AVAILABLE FROM THE PROTEIN DATA REMARK 8 BANK AS ENTRY 1CCP AND THREE HEME-CLEFT MUTANTS PREPARED BY REMARK 8 SITE-DIRECTED MUTAGENESIS, CCP(MI,D235N), CCP(MI,W191F), REMARK 8 AND CCP(MI,W51F), ARE ALSO AVAILABLE FROM THE PROTEIN DATA REMARK 8 BANK AS ENTRIES 2CCP, 3CCP, AND 4CCP, RESPECTIVELY. REMARK 8 OTHER MUTANT STRUCTURES - CCP(MI,H52L), AND CCP(MI,R48K) REMARK 8 HAVE RECENTLY BEEN DEPOSITED AS 5CCP AND 7CCP, REMARK 8 RESPECTIVELY. REMARK 9 REMARK 9 ACTIVE SITE WATER MOLECULE HOH-595 HAD VERY LOW OCCUPANCY REMARK 9 IN THE R48L MUTANT, AND WAS NOT INCLUDED IN THE MODEL. REMARK 10 REMARK 10 SOME ATOMS IN THE FOLLOWING RESIDUES COULD NOT BE LOCATED; REMARK 10 LYS 12, GLU 35, LYS 74, LYS 90, LYS 97, LYS 183, LYS 226, REMARK 10 LYS 260, LYS 278. SEQRES 1 296 MET ILE THR THR PRO LEU VAL HIS VAL ALA SER VAL GLU SEQRES 2 296 LYS GLY ARG SER TYR GLU ASP PHE GLN LYS VAL TYR ASN SEQRES 3 296 ALA ILE ALA LEU LYS LEU ARG GLU ASP ASP GLU TYR ASP SEQRES 4 296 ASN TYR ILE GLY TYR GLY PRO VAL LEU VAL LEU LEU ALA SEQRES 5 296 TRP HIS ILE SER GLY THR TRP ASP LYS HIS ASP ASN THR SEQRES 6 296 GLY GLY SER TYR GLY GLY THR TYR ARG PHE LYS LYS GLU SEQRES 7 296 PHE ASN ASP PRO SER ASN ALA GLY LEU GLN ASN GLY PHE SEQRES 8 296 LYS PHE LEU GLU PRO ILE HIS LYS GLU PHE PRO TRP ILE SEQRES 9 296 SER SER GLY ASP LEU PHE SER LEU GLY GLY VAL THR ALA SEQRES 10 296 VAL GLN GLU MET GLN GLY PRO LYS ILE PRO TRP ARG CYS SEQRES 11 296 GLY ARG VAL ASP THR PRO GLU ASP THR THR PRO ASP ASN SEQRES 12 296 GLY ARG LEU PRO ASP ALA ASP LYS ASP ALA GLY TYR VAL SEQRES 13 296 ARG THR PHE PHE GLN ARG LEU ASN MET ASN ASP ARG GLU SEQRES 14 296 VAL VAL ALA LEU MET GLY ALA HIS ALA LEU GLY LYS THR SEQRES 15 296 HIS LEU LYS ASN SER GLY TYR GLU GLY PRO TRP GLY ALA SEQRES 16 296 ALA ASN ASN VAL PHE THR ASN GLU PHE TYR LEU ASN LEU SEQRES 17 296 LEU ASN GLU ASP TRP LYS LEU GLU LYS ASN ASP ALA ASN SEQRES 18 296 ASN GLU GLN TRP ASP SER LYS SER GLY TYR MET MET LEU SEQRES 19 296 PRO THR ASP TYR SER LEU ILE GLN ASP PRO LYS TYR LEU SEQRES 20 296 SER ILE VAL LYS GLU TYR ALA ASN ASP GLN ASP LYS PHE SEQRES 21 296 PHE LYS ASP PHE SER LYS ALA PHE GLU LYS LEU LEU GLU SEQRES 22 296 ASN GLY ILE THR PHE PRO LYS ASP ALA PRO SER PRO PHE SEQRES 23 296 ILE PHE LYS THR LEU GLU GLU GLN GLY LEU HET HEM 296 43 PROTOPORPHYRIN IX CONTAINS FE(III) FORMUL 2 HEM C34 H32 N4 O4 FE1 +++ FORMUL 3 HOH *210(H2 O1) HELIX 1 A SER 15 ASP 33 1 HELIX 2 B TYR 42 SER 54 1 HELIX 3 B1 PHE 73 ASN 78 1 HELIX 4 C GLY 84 PHE 99 1 BENT AT PRO 94 HELIX 5 D SER 103 MET 119 1 HELIX 6 E ASP 150 PHE 158 1 HELIX 7 F ASN 164 LEU 177 1 HELIX 8 F1 HIS 181 GLY 186 1 HELIX 9 G GLU 201 GLU 209 1 HELIX 10 H LEU 232 GLN 240 1 HELIX 11 I ASP 241 ASN 253 1 HELIX 12 J GLN 255 ASN 272 1 HELIX 13 J1 THR 288 GLY 293 1 TURN 1 T1 GLU 11 ARG 14 TYPE II TURN 2 T2 ASP 37 ILE 40 TYPE I' TURN 3 T3 ASN 82 LEU 85 TYPE II TURN 4 T4 ASP 58 ASP 61 TYPE I TURN 5 T5 ASN 216 ASN 219 TYPE I TURN 6 T6 SER 225 GLY 228 TYPE I TURN 7 T7 GLU 271 ILE 274 TYPE II CRYST1 105.200 74.280 45.110 90.00 90.00 90.00 P 21 21 21 4 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 0.009506 0.000000 0.000000 0.00000 SCALE2 0.000000 0.013463 0.000000 0.00000 SCALE3 0.000000 0.000000 0.022168 0.00000