HEADER OXIDOREDUCTASE(OXYGENASE) 16-SEP-93 2HPD COMPND CYTOCHROME P450 (BM-3) (E.C.1.14.14.1) (HEMOPROTEIN DOMAIN) COMPND 2 (FATTY ACID MONOOXYGENASE) SOURCE (BACILLUS MEGATERIUM) RECOMBINANT FORM EXPRESSED SOURCE 2 IN (ESCHERICHIA COLI) AUTHOR K.G.RAVICHANDRAN,S.S.BODDUPALLI,C.A.HASEMANN,J.A.PETERSON, AUTHOR 2 J.DEISENHOFER REVDAT 1 31-OCT-93 2HPD 0 JRNL AUTH K.G.RAVICHANDRAN,S.S.BODDUPALLI,C.A.HASEMANN, JRNL AUTH 2 J.A.PETERSON,J.DEISENHOFER JRNL TITL CRYSTAL STRUCTURE OF HEMOPROTEIN DOMAIN OF JRNL TITL 2 P450BM-3, A PROTOTYPE FOR MICROSOMAL P450'S JRNL REF SCIENCE V. 261 731 1993 JRNL REFN ASTM SCIEAS US ISSN 0036-8075 038 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH S.S.BODDUPALLI,C.A.HASEMANN,K.G.RAVICHANDRAN, REMARK 1 AUTH 2 J.-Y.LU,E.J.GOLDSMITH,J.DEISENHOFER,J.A.PETERSON REMARK 1 TITL CRYSTALLIZATION AND PRELIMINARY X-RAY DIFFRACTION REMARK 1 TITL 2 ANALYSIS OF P450=TERP= AND THE HEMOPROTEIN DOMAIN REMARK 1 TITL 3 OF P450=BM-3=, ENZYMES BELONGING TO TWO DISTINCT REMARK 1 TITL 4 CLASSES OF THE CYTOCHROME P450 SUPERFAMILY REMARK 1 REF PROC.NAT.ACAD.SCI.USA V. 89 5567 1992 REMARK 1 REFN ASTM PNASA6 US ISSN 0027-8424 040 REMARK 1 REFERENCE 2 REMARK 1 AUTH A.J.FULCO REMARK 1 TITL P450=BM-3= AND OTHER INDUCIBLE BACTERIAL P450 REMARK 1 TITL 2 CYTOCHROMES: BIOCHEMISTRY AND REGULATION REMARK 1 REF ANNU.REV.PHARMACOL.TOXICOL. V. 31 177 1991 REMARK 1 REFN ASTM ARPTDI US ISSN 0362-1642 756 REMARK 2 REMARK 2 RESOLUTION. 2.0 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM X-PLOR REMARK 3 AUTHORS BRUNGER REMARK 3 R VALUE 0.167 REMARK 3 RMSD BOND DISTANCES 0.016 ANGSTROMS REMARK 3 RMSD BOND ANGLES 3.1 DEGREES REMARK 3 REMARK 3 RESOLUTION RANGE 20.0 - 2.0 ANGSTROMS REMARK 4 REMARK 4 THE TWO MOLECULES IN THE ASYMMETRIC UNIT HAVE BEEN ASSIGNED REMARK 4 CHAIN INDICATORS *A* AND *B*. THIS ENTRY INCLUDES RESIDUES REMARK 4 1 - 457 AND THE HEME FOR EACH MOLECULE. THE FOURTEEN REMARK 4 C-TERMINAL RESIDUES OF THE POLYPEPTIDE ARE DISORDERED IN REMARK 4 THE CRYSTALS FOR BOTH MOLECULES. REMARK 5 REMARK 5 THE TRANSFORMATION PRESENTED ON *MTRIX 1* RECORDS BELOW REMARK 5 WILL YIELD APPROXIMATE COORDINATES FOR CHAIN *B* WHEN REMARK 5 APPLIED TO CHAIN *A*, WITH AN RMSD = 0.779480. THE REMARK 5 TRANSFORMATION PRESENTED ON *MTRIX 2* RECORDS BELOW WILL REMARK 5 YIELD APPROXIMATE COORDINATES FOR CHAIN *A* WHEN APPLIED TO REMARK 5 CHAIN *B*, WITH AN RMSD = 0.785550. REMARK 6 REMARK 6 THE ELECTRON DENSITY FOR THE LOOP REGION BETWEEN THE F AND REMARK 6 G HELICES (RESIDUES 190 - 196) IS POORLY DEFINED FOR BOTH REMARK 6 MOLECULES A AND B. THIS COULD ACCOUNT FOR THE HIGH B REMARK 6 FACTORS OF RESIDUES IN THIS REGION AND FOR THE BAD REMARK 6 CONFORMATION OF THE 195 - 196 PEPTIDE PLANE. REMARK 7 REMARK 7 DIHEDRAL ANGLES OF RESIDUES THR A 436 AND THR B 436 LIE REMARK 7 OUTSIDE THE ALLOWED REGIONS IN THE RAMACHANDRAN PLOT. THIS REMARK 7 MAY BE DUE TO THEIR ROLE IN SUBSTRATE INTERACTIONS. REMARK 8 REMARK 8 THE HEME IRON IS COVALENTLY BONDED TO AN OXYGEN OF WATER. SEQRES 1 A 471 THR ILE LYS GLU MET PRO GLN PRO LYS THR PHE GLY GLU SEQRES 2 A 471 LEU LYS ASN LEU PRO LEU LEU ASN THR ASP LYS PRO VAL SEQRES 3 A 471 GLN ALA LEU MET LYS ILE ALA ASP GLU LEU GLY GLU ILE SEQRES 4 A 471 PHE LYS PHE GLU ALA PRO GLY ARG VAL THR ARG TYR LEU SEQRES 5 A 471 SER SER GLN ARG LEU ILE LYS GLU ALA CYS ASP GLU SER SEQRES 6 A 471 ARG PHE ASP LYS ASN LEU SER GLN ALA LEU LYS PHE VAL SEQRES 7 A 471 ARG ASP PHE ALA GLY ASP GLY LEU PHE THR SER TRP THR SEQRES 8 A 471 HIS GLU LYS ASN TRP LYS LYS ALA HIS ASN ILE LEU LEU SEQRES 9 A 471 PRO SER PHE SER GLN GLN ALA MET LYS GLY TYR HIS ALA SEQRES 10 A 471 MET MET VAL ASP ILE ALA VAL GLN LEU VAL GLN LYS TRP SEQRES 11 A 471 GLU ARG LEU ASN ALA ASP GLU HIS ILE GLU VAL PRO GLU SEQRES 12 A 471 ASP MET THR ARG LEU THR LEU ASP THR ILE GLY LEU CYS SEQRES 13 A 471 GLY PHE ASN TYR ARG PHE ASN SER PHE TYR ARG ASP GLN SEQRES 14 A 471 PRO HIS PRO PHE ILE THR SER MET VAL ARG ALA LEU ASP SEQRES 15 A 471 GLU ALA MET ASN LYS LEU GLN ARG ALA ASN PRO ASP ASP SEQRES 16 A 471 PRO ALA TYR ASP GLU ASN LYS ARG GLN PHE GLN GLU ASP SEQRES 17 A 471 ILE LYS VAL MET ASN ASP LEU VAL ASP LYS ILE ILE ALA SEQRES 18 A 471 ASP ARG LYS ALA SER GLY GLU GLN SER ASP ASP LEU LEU SEQRES 19 A 471 THR HIS MET LEU ASN GLY LYS ASP PRO GLU THR GLY GLU SEQRES 20 A 471 PRO LEU ASP ASP GLU ASN ILE ARG TYR GLN ILE ILE THR SEQRES 21 A 471 PHE LEU ILE ALA GLY HIS GLU THR THR SER GLY LEU LEU SEQRES 22 A 471 SER PHE ALA LEU TYR PHE LEU VAL LYS ASN PRO HIS VAL SEQRES 23 A 471 LEU GLN LYS ALA ALA GLU GLU ALA ALA ARG VAL LEU VAL SEQRES 24 A 471 ASP PRO VAL PRO SER TYR LYS GLN VAL LYS GLN LEU LYS SEQRES 25 A 471 TYR VAL GLY MET VAL LEU ASN GLU ALA LEU ARG LEU TRP SEQRES 26 A 471 PRO THR ALA PRO ALA PHE SER LEU TYR ALA LYS GLU ASP SEQRES 27 A 471 THR VAL LEU GLY GLY GLU TYR PRO LEU GLU LYS GLY ASP SEQRES 28 A 471 GLU LEU MET VAL LEU ILE PRO GLN LEU HIS ARG ASP LYS SEQRES 29 A 471 THR ILE TRP GLY ASP ASP VAL GLU GLU PHE ARG PRO GLU SEQRES 30 A 471 ARG PHE GLU ASN PRO SER ALA ILE PRO GLN HIS ALA PHE SEQRES 31 A 471 LYS PRO PHE GLY ASN GLY GLN ARG ALA CYS ILE GLY GLN SEQRES 32 A 471 GLN PHE ALA LEU HIS GLU ALA THR LEU VAL LEU GLY MET SEQRES 33 A 471 MET LEU LYS HIS PHE ASP PHE GLU ASP HIS THR ASN TYR SEQRES 34 A 471 GLU LEU ASP ILE LYS GLU THR LEU THR LEU LYS PRO GLU SEQRES 35 A 471 GLY PHE VAL VAL LYS ALA LYS SER LYS LYS ILE PRO LEU SEQRES 36 A 471 GLY GLY ILE PRO SER PRO SER THR GLU GLN SER ALA LYS SEQRES 37 A 471 LYS VAL ARG SEQRES 1 B 471 THR ILE LYS GLU MET PRO GLN PRO LYS THR PHE GLY GLU SEQRES 2 B 471 LEU LYS ASN LEU PRO LEU LEU ASN THR ASP LYS PRO VAL SEQRES 3 B 471 GLN ALA LEU MET LYS ILE ALA ASP GLU LEU GLY GLU ILE SEQRES 4 B 471 PHE LYS PHE GLU ALA PRO GLY ARG VAL THR ARG TYR LEU SEQRES 5 B 471 SER SER GLN ARG LEU ILE LYS GLU ALA CYS ASP GLU SER SEQRES 6 B 471 ARG PHE ASP LYS ASN LEU SER GLN ALA LEU LYS PHE VAL SEQRES 7 B 471 ARG ASP PHE ALA GLY ASP GLY LEU PHE THR SER TRP THR SEQRES 8 B 471 HIS GLU LYS ASN TRP LYS LYS ALA HIS ASN ILE LEU LEU SEQRES 9 B 471 PRO SER PHE SER GLN GLN ALA MET LYS GLY TYR HIS ALA SEQRES 10 B 471 MET MET VAL ASP ILE ALA VAL GLN LEU VAL GLN LYS TRP SEQRES 11 B 471 GLU ARG LEU ASN ALA ASP GLU HIS ILE GLU VAL PRO GLU SEQRES 12 B 471 ASP MET THR ARG LEU THR LEU ASP THR ILE GLY LEU CYS SEQRES 13 B 471 GLY PHE ASN TYR ARG PHE ASN SER PHE TYR ARG ASP GLN SEQRES 14 B 471 PRO HIS PRO PHE ILE THR SER MET VAL ARG ALA LEU ASP SEQRES 15 B 471 GLU ALA MET ASN LYS LEU GLN ARG ALA ASN PRO ASP ASP SEQRES 16 B 471 PRO ALA TYR ASP GLU ASN LYS ARG GLN PHE GLN GLU ASP SEQRES 17 B 471 ILE LYS VAL MET ASN ASP LEU VAL ASP LYS ILE ILE ALA SEQRES 18 B 471 ASP ARG LYS ALA SER GLY GLU GLN SER ASP ASP LEU LEU SEQRES 19 B 471 THR HIS MET LEU ASN GLY LYS ASP PRO GLU THR GLY GLU SEQRES 20 B 471 PRO LEU ASP ASP GLU ASN ILE ARG TYR GLN ILE ILE THR SEQRES 21 B 471 PHE LEU ILE ALA GLY HIS GLU THR THR SER GLY LEU LEU SEQRES 22 B 471 SER PHE ALA LEU TYR PHE LEU VAL LYS ASN PRO HIS VAL SEQRES 23 B 471 LEU GLN LYS ALA ALA GLU GLU ALA ALA ARG VAL LEU VAL SEQRES 24 B 471 ASP PRO VAL PRO SER TYR LYS GLN VAL LYS GLN LEU LYS SEQRES 25 B 471 TYR VAL GLY MET VAL LEU ASN GLU ALA LEU ARG LEU TRP SEQRES 26 B 471 PRO THR ALA PRO ALA PHE SER LEU TYR ALA LYS GLU ASP SEQRES 27 B 471 THR VAL LEU GLY GLY GLU TYR PRO LEU GLU LYS GLY ASP SEQRES 28 B 471 GLU LEU MET VAL LEU ILE PRO GLN LEU HIS ARG ASP LYS SEQRES 29 B 471 THR ILE TRP GLY ASP ASP VAL GLU GLU PHE ARG PRO GLU SEQRES 30 B 471 ARG PHE GLU ASN PRO SER ALA ILE PRO GLN HIS ALA PHE SEQRES 31 B 471 LYS PRO PHE GLY ASN GLY GLN ARG ALA CYS ILE GLY GLN SEQRES 32 B 471 GLN PHE ALA LEU HIS GLU ALA THR LEU VAL LEU GLY MET SEQRES 33 B 471 MET LEU LYS HIS PHE ASP PHE GLU ASP HIS THR ASN TYR SEQRES 34 B 471 GLU LEU ASP ILE LYS GLU THR LEU THR LEU LYS PRO GLU SEQRES 35 B 471 GLY PHE VAL VAL LYS ALA LYS SER LYS LYS ILE PRO LEU SEQRES 36 B 471 GLY GLY ILE PRO SER PRO SER THR GLU GLN SER ALA LYS SEQRES 37 B 471 LYS VAL ARG FTNOTE 1 FTNOTE 1 RESIDUES ALA 191 - ALA 197 OF BOTH MOLECULES HAVE LARGE B FTNOTE 1 VALUES (B > 80 ANGSTROMS**2). FTNOTE 2 FTNOTE 2 ASP A 195 - PRO A 196 OMEGA =104.96 FTNOTE 2 PEPTIDE BOND DEVIATES SIGNIFICANTLY FROM TRANS CONFORMATION FTNOTE 2 FTNOTE 1 ALSO APPLIES. HET HEM A 472 44 PROTOPORPHYRIN IX CONTAINS FE(III) AND O HET HEM B 472 44 PROTOPORPHYRIN IX CONTAINS FE(III) AND O FORMUL 3 HEM 2(C34 H32 N4 O4 FE1 +++). FORMUL 3 HEM 2(O1) FORMUL 4 HOH *453(H2 O1) HELIX 1 AA ASN A 16 LEU A 20 5 HELIX 2 A1A PRO A 25 GLY A 37 1 HELIX 3 BA SER A 54 CYS A 62 1 HELIX 4 B'A SER A 72 GLY A 83 1 HELIX 5 CA GLU A 93 LEU A 104 1 HELIX 6 B1A PRO A 105 SER A 108 5 HELIX 7 C1A GLN A 109 LYS A 113 5 HELIX 8 DA GLY A 114 ARG A 132 1 HELIX 9 EA PRO A 142 PHE A 158 1 HELIX 10 D1A ASN A 163 ARG A 167 5 HELIX 11 FA PRO A 172 GLN A 189 1 HELIX 12 GA ALA A 197 SER A 226 1 HELIX 13 HA LEU A 233 ASN A 239 1 HELIX 14 IA ASP A 250 LYS A 282 1 HELIX 15 JA ASN A 283 LEU A 298 1 HELIX 16 J'A SER A 304 GLN A 310 1 HELIX 17 KA LEU A 311 TRP A 325 1 HELIX 18 K'A ILE A 357 HIS A 361 1 HELIX 19 E1A ARG A 375 PHE A 379 5 HELIX 20 F1A ASN A 381 ILE A 385 5 HELIX 21 LA GLY A 402 LYS A 419 1 HELIX 22 AB ASN B 16 LEU B 20 5 HELIX 23 A1B PRO B 25 GLY B 37 1 HELIX 24 BB SER B 54 CYS B 62 1 HELIX 25 B'B SER B 72 GLY B 83 1 HELIX 26 CB GLU B 93 LEU B 104 1 HELIX 27 B1B PRO B 105 SER B 108 5 HELIX 28 C1B GLN B 109 LYS B 113 5 HELIX 29 DB GLY B 114 ARG B 132 1 HELIX 30 EB PRO B 142 PHE B 158 1 HELIX 31 D1B ASN B 163 ARG B 167 5 HELIX 32 FB PRO B 172 GLN B 189 1 HELIX 33 GB ALA B 197 SER B 226 1 HELIX 34 HB LEU B 233 ASN B 239 1 HELIX 35 IB ASP B 250 LYS B 282 1 HELIX 36 JB ASN B 283 LEU B 298 1 HELIX 37 J'B SER B 304 GLN B 310 1 HELIX 38 KB LEU B 311 TRP B 325 1 HELIX 39 K'B ILE B 357 HIS B 361 1 HELIX 40 E1B ARG B 375 PHE B 379 5 HELIX 41 F1B ASN B 381 ILE B 385 5 HELIX 42 LB GLY B 402 LYS B 419 1 SHEET 1 1A 5 GLU A 38 ALA A 44 0 SHEET 2 1A 5 ARG A 47 SER A 53 -1 SHEET 3 1A 5 GLY A 350 LEU A 356 1 SHEET 4 1A 5 PRO A 329 ALA A 335 -1 SHEET 5 1A 5 ARG A 66 ASN A 70 -1 SHEET 1 2A 2 GLU A 337 LEU A 341 0 SHEET 2 2A 2 GLU A 344 GLU A 348 -1 SHEET 1 3A 3 HIS A 138 VAL A 141 0 SHEET 2 3A 3 PHE A 444 LYS A 451 -1 SHEET 3 3A 3 HIS A 420 GLU A 424 -1 SHEET 1 4A 2 ASP A 432 THR A 436 0 SHEET 2 4A 2 THR A 438 GLU A 442 -1 SHEET 1 1B 5 GLU B 38 ALA B 44 0 SHEET 2 1B 5 ARG B 47 SER B 53 -1 SHEET 3 1B 5 GLY B 350 LEU B 356 1 SHEET 4 1B 5 PRO B 329 ALA B 335 -1 SHEET 5 1B 5 ARG B 66 ASN B 70 -1 SHEET 1 2B 2 GLU B 337 LEU B 341 0 SHEET 2 2B 2 GLU B 344 GLU B 348 -1 SHEET 1 3B 3 HIS B 138 VAL B 141 0 SHEET 2 3B 3 PHE B 444 LYS B 451 -1 SHEET 3 3B 3 HIS B 420 GLU B 424 -1 SHEET 1 4B 2 ASP B 432 THR B 436 0 SHEET 2 4B 2 THR B 438 GLU B 442 -1 CRYST1 59.400 154.000 62.200 90.00 94.70 90.00 P 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 0.016835 0.000000 0.001384 0.00000 SCALE2 0.000000 0.006494 0.000000 0.00000 SCALE3 0.000000 0.000000 0.016131 0.00000 MTRIX1 1 0.999600 0.016600 0.021100 -23.70000 1 MTRIX2 1 0.008100 -0.934400 0.356200 39.90000 1 MTRIX3 1 0.025700 -0.355900 -0.934200 21.20000 1 MTRIX1 2 0.999600 0.008100 0.025700 22.80000 1 MTRIX2 2 0.016700 -0.934400 -0.355900 45.20000 1 MTRIX3 2 0.021100 0.356200 -0.934200 6.18000 1