HEADER OXIDOREDUCTASE(OXYGEN RECEPTOR) 14-JUN-93 3COX COMPND CHOLESTEROL OXIDASE (E.C.1.1.3.6) SOURCE (BREVIBACTERIUM STEROLICUM) AUTHOR A.VRIELINK,J.LI,P.BRICK,D.M.BLOW REVDAT 1 31-OCT-93 3COX 0 SPRSDE 15-OCT-93 3COX 1COX JRNL AUTH J.LI,A.VRIELINK,P.BRICK,D.M.BLOW JRNL TITL CRYSTAL STRUCTURE OF CHOLESTEROL OXIDASE JRNL TITL 2 COMPLEXED WITH A STEROID SUBSTRATE. IMPLICATIONS JRNL TITL 3 FOR FAD DEPENDENT ALCOHOL OXIDASES JRNL REF TO BE PUBLISHED JRNL REFN 353 REMARK 1 REMARK 1 REFERENCE 1 REMARK 1 AUTH A.VRIELINK,L.F.LLOYD,D.M.BLOW REMARK 1 TITL CRYSTAL STRUCTURE OF CHOLESTEROL OXIDASE FROM REMARK 1 TITL 2 BREVIBACTERIUM STEROLICUM REFINED AT 1.8 REMARK 1 TITL 3 ANGSTROMS RESOLUTION REMARK 1 REF J.MOL.BIOL. V. 219 533 1991 REMARK 1 REFN ASTM JMOBAK UK ISSN 0022-2836 070 REMARK 1 REFERENCE 2 REMARK 1 AUTH T.OHTA,K.FUJISHIRO,K.YAMAGUCHI,Y.TAMURA,K.AISAKA, REMARK 1 AUTH 2 T.UWAJIMA,M.HASEGAWA REMARK 1 TITL SEQUENCE OF GENE CHOB ENCODING CHOLESTEROL REMARK 1 TITL 2 OXIDASE OF BREVIBACTERIUM STEROLICUM: COMPARISON REMARK 1 TITL 3 WITH CHOA OF STREPTOMYCES SP. SA-COO REMARK 1 REF GENE V. 103 93 1991 REMARK 1 REFN ASTM GENED6 NE ISSN 0378-1119 861 REMARK 2 REMARK 2 RESOLUTION. 1.8 ANGSTROMS. REMARK 3 REMARK 3 REFINEMENT. REMARK 3 PROGRAM X-PLOR REMARK 3 AUTHORS BRUNGER REMARK 3 R VALUE 0.156 REMARK 3 RMSD BOND DISTANCES 0.011 ANGSTROMS REMARK 3 REMARK 3 RESOLUTION RANGE 10.0 - 1.8 ANGSTROMS REMARK 3 DATA CUTOFF 0.0 SIGMA(F) REMARK 3 REMARK 3 NUMBER OF PROTEIN ATOMS 3791 REMARK 3 NUMBER OF SOLVENT ATOMS 453 REMARK 3 REMARK 3 THE GEOMETRIC RESTRAINTS USED FOR THE PROTEIN WERE BASE REMARK 3 ON THOSE PUBLISHED BY R.A. ENGH AND R. HUBER (ACTA CRYST. REMARK 3 A47, 392-400 (1991)) AS DISTRIBUTED WITH VERSION 3.1 OF REMARK 3 *X-PLOR*. THE BOND LENGTH AND BOND ANGLE PARAMETERS OF REMARK 3 THE FLAVIN SYSTEM WERE OBTAINED FROM SMALL MOLECULE REMARK 3 CRYSTAL STRUCTURES DEPOSITED IN THE CAMBRIDGE STRUCTURAL REMARK 3 DATABASE. A LOWER LIMIT OF 2.0 ANGSTROM**2 HAS BEEN USED REMARK 3 FOR THE ATOMIC TEMPERATURE FACTORS. REMARK 4 REMARK 4 RESIDUE 437 HAS BEEN BUILT AS A TYROSINE BUT IS GIVEN AS A REMARK 4 PHENYLALANINE IN REFERENCE 2. THE HYDROXYL GROUP HAS REMARK 4 SIGNIFICANT ELECTRON DENSITY. REMARK 5 REMARK 5 RESIDUE 492 HAS BEEN BUILT AS AN ALANINE BUT IS GIVEN AS AN REMARK 5 ARGININE IN REFERENCE 2. A CONFORMATIONAL CHANGE IN THE REMARK 5 PROTEIN STRUCTURE WOULD BE NECESSARY TO ACCOMMODATE AN REMARK 5 ARGININE AT THIS POSITION. REMARK 6 REMARK 6 THE FOLLOWING RESIDUES HAVE NO SIGNIFICANT ELECTRON DENSITY REMARK 6 IN FINAL MAP AND HAVE CONSEQUENTLY BEEN OMITTED FROM THE REMARK 6 MODEL: 1 - 4, 434 - 435, 507. REMARK 6 REMARK 7 THE FOLLOWING RESIDUES CONTAIN SIDE-CHAIN ATOMS WHICH HAVE REMARK 7 NO SIGNIFICANT ELECTRON DENSITY IN THE FINAL MAP AND HAVE REMARK 7 THEREFORE BEEN OMITTED FROM THE MODEL: 11, 51, 54, 61, 64, REMARK 7 71, 72, 79, 85, 87, 91, 100, 105, 169, 202, 207, 230, 339, REMARK 7 387, 396, 414, 415, 419, 422, 436, 438, 500. REMARK 8 REMARK 8 THE STRUCTURE CONTAINS AN FAD COFACTOR IN THE OXIDIZED REMARK 8 FORM. THE FLAVIN RING SYSTEM OF THE FAD MOLECULE ADOPTS A REMARK 8 SLIGHTLY TWISTED CONFORMATION. SEQRES 1 507 ALA PRO SER ARG THR LEU ALA ASP GLY ASP ARG VAL PRO SEQRES 2 507 ALA LEU VAL ILE GLY SER GLY TYR GLY GLY ALA VAL ALA SEQRES 3 507 ALA LEU ARG LEU THR GLN ALA GLY ILE PRO THR GLN ILE SEQRES 4 507 VAL GLU MET GLY ARG SER TRP ASP THR PRO GLY SER ASP SEQRES 5 507 GLY LYS ILE PHE CYS GLY MET LEU ASN PRO ASP LYS ARG SEQRES 6 507 SER MET TRP LEU ALA ASP LYS THR ASP GLN PRO VAL SER SEQRES 7 507 ASN PHE MET GLY PHE GLY ILE ASN LYS SER ILE ASP ARG SEQRES 8 507 TYR VAL GLY VAL LEU ASP SER GLU ARG PHE SER GLY ILE SEQRES 9 507 LYS VAL TYR GLN GLY ARG GLY VAL GLY GLY GLY SER LEU SEQRES 10 507 VAL ASN GLY GLY MET ALA VAL THR PRO LYS ARG ASN TYR SEQRES 11 507 PHE GLU GLU ILE LEU PRO SER VAL ASP SER ASN GLU MET SEQRES 12 507 TYR ASN LYS TYR PHE PRO ARG ALA ASN THR GLY LEU GLY SEQRES 13 507 VAL ASN ASN ILE ASP GLN ALA TRP PHE GLU SER THR GLU SEQRES 14 507 TRP TYR LYS PHE ALA ARG THR GLY ARG LYS THR ALA GLN SEQRES 15 507 ARG SER GLY PHE THR THR ALA PHE VAL PRO ASN VAL TYR SEQRES 16 507 ASP PHE GLU TYR MET LYS LYS GLU ALA ALA GLY GLN VAL SEQRES 17 507 THR LYS SER GLY LEU GLY GLY GLU VAL ILE TYR GLY ASN SEQRES 18 507 ASN ALA GLY LYS LYS SER LEU ASP LYS THR TYR LEU ALA SEQRES 19 507 GLN ALA ALA ALA THR GLY LYS LEU THR ILE THR THR LEU SEQRES 20 507 HIS ARG VAL THR LYS VAL ALA PRO ALA THR GLY SER GLY SEQRES 21 507 TYR SER VAL THR MET GLU GLN ILE ASP GLU GLN GLY ASN SEQRES 22 507 VAL VAL ALA THR LYS VAL VAL THR ALA ASP ARG VAL PHE SEQRES 23 507 PHE ALA ALA GLY SER VAL GLY THR SER LYS LEU LEU VAL SEQRES 24 507 SER MET LYS ALA GLN GLY HIS LEU PRO ASN LEU SER SER SEQRES 25 507 GLN VAL GLY GLU GLY TRP GLY ASN ASN GLY ASN ILE MET SEQRES 26 507 VAL GLY ARG ALA ASN HIS MET TRP ASP ALA THR GLY SER SEQRES 27 507 LYS GLN ALA THR ILE PRO THR MET GLY ILE ASP ASN TRP SEQRES 28 507 ALA ASP PRO THR ALA PRO ILE PHE ALA GLU ILE ALA PRO SEQRES 29 507 LEU PRO ALA GLY LEU GLU THR TYR VAL SER LEU TYR LEU SEQRES 30 507 ALA ILE THR LYS ASN PRO GLU ARG ALA ARG PHE GLN PHE SEQRES 31 507 ASN SER GLY THR GLY LYS VAL ASP LEU THR TRP ALA GLN SEQRES 32 507 SER GLN ASN GLN LYS GLY ILE ASP MET ALA LYS LYS VAL SEQRES 33 507 PHE ASP LYS ILE ASN GLN LYS GLU GLY THR ILE TYR ARG SEQRES 34 507 THR ASP LEU PHE GLY VAL TYR TYR LYS THR TRP GLY ASP SEQRES 35 507 ASP PHE THR TYR HIS PRO LEU GLY GLY VAL LEU LEU ASN SEQRES 36 507 LYS ALA THR ASP ASN PHE GLY ARG LEU PRO GLU TYR PRO SEQRES 37 507 GLY LEU TYR VAL VAL ASP GLY SER LEU VAL PRO GLY ASN SEQRES 38 507 VAL GLY VAL ASN PRO PHE VAL THR ILE THR ALA LEU ALA SEQRES 39 507 GLU ARG ASN MET ASP LYS ILE ILE SER SER ASP ILE GLN HET FAD 510 53 FLAVIN-ADENINE DINUCLEOTIDE FORMUL 2 FAD C27 H33 N9 O15 P2 FORMUL 3 HOH *453(H2 O1) HELIX 1 H1 TYR 21 GLN 32 1 HELIX 2 H2 ARG 128 ILE 134 1 HELIX 3 H3A SER 140 ASN 145 1 HELIX 4 H3B TYR 147 LEU 155 1 HELIX 5 H4 GLN 162 SER 167 1 HELIX 6 H5A GLU 169 TYR 171 5 HELIX 7 H5B LYS 172 ARG 183 1 HELIX 8 H6 PHE 197 ALA 204 1 HELIX 9 H7A LEU 228 LYS 230 5 HELIX 10 H7B TYR 232 ALA 238 1 HELIX 11 H8 GLY 290 ALA 303 1 HELIX 12 H9A GLN 403 ASN 406 5 HELIX 13 H9B GLN 407 GLU 424 1 HELIX 14 H10 PHE 487 SER 504 1 SHEET 1 A 6 LEU 242 THR 245 0 SHEET 2 A 6 THR 37 VAL 40 1 O THR 37 N THR 243 SHEET 3 A 6 ALA 14 ILE 17 1 O ALA 14 N GLN 38 SHEET 4 A 6 ARG 284 PHE 287 1 O ARG 284 N LEU 15 SHEET 5 A 6 LEU 470 VAL 472 1 O TYR 471 N PHE 287 SHEET 6 A 6 ARG 463 PRO 465 -1 N LEU 464 O LEU 470 SHEET 1 B 6 THR 188 PHE 190 0 SHEET 2 B 6 MET 346 ASP 349 -1 N GLY 347 O ALA 189 SHEET 3 B 6 ILE 358 ILE 362 -1 N ALA 360 O ILE 348 SHEET 4 B 6 VAL 373 THR 380 -1 N ILE 379 O PHE 359 SHEET 5 B 6 ILE 324 ALA 329 -1 N VAL 326 O TYR 376 SHEET 6 B 6 THR 439 GLY 441 -1 N GLY 441 O MET 325 SHEET 1 C 4 ARG 11 VAL 12 0 SHEET 2 C 4 VAL 274 ALA 282 1 O THR 281 N VAL 12 SHEET 3 C 4 TYR 261 ILE 268 -1 O TYR 261 N ALA 282 SHEET 4 C 4 HIS 248 PRO 255 -1 N ARG 249 O GLU 266 SHEET 1 D 3 LEU 96 ARG 100 0 SHEET 2 D 3 LYS 105 GLY 109 -1 O GLN 108 N ASP 97 SHEET 3 D 3 PHE 444 THR 445 1 N THR 445 O LYS 105 SHEET 1 E 2 TRP 318 GLY 319 0 SHEET 2 E 2 PRO 448 LEU 449 -1 O LEU 449 N GLY 319 SHEET 1 F 2 PHE 388 ASN 391 0 SHEET 2 F 2 LYS 396 LEU 399 -1 N ASP 398 O GLN 389 TURN 1 T1 ALA 7 ASP 10 TURN 2 T2 GLN 32 ILE 35 TURN 3 T3 GLY 50 GLY 53 TURN 4 T4 ASP 63 SER 66 TURN 5 T5 PHE 80 PHE 83 TURN 6 T6 GLY 111 GLY 114 BURIED TURN TURN 7 T7 GLY 113 SER 116 TURN 8 T8 LEU 135 VAL 138 TURN 9 T9 SER 211 GLY 214 TURN 10 T10 VAL 217 GLY 220 BURIED TURN 11 T11 SER 227 LYS 230 TURN 12 T12 ASP 269 GLY 272 TURN 13 T13 LEU 307 LEU 310 TURN 14 T14 GLN 313 GLU 316 TURN 15 T15 ASN 320 ASN 323 BURIED TURN TURN 16 T16 HIS 331 ASP 334 TURN 17 T17 ASN 350 ASP 353 TURN 18 T18 ASP 353 ALA 356 TURN 19 T19 LEU 453 LYS 456 TURN 20 T20 ASP 459 GLY 462 TURN 21 T21 LEU 464 TYR 467 TURN 22 T22 TYR 467 LEU 470 TURN 23 T23 ASP 474 LEU 477 BURIED TURN CRYST1 67.720 84.580 88.130 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 0.014767 0.000000 0.000000 0.00000 SCALE2 0.000000 0.011823 0.000000 0.00000 SCALE3 0.000000 0.000000 0.011347 0.00000