ID CHOD_STRS0 Reviewed; 546 AA. AC P12676; DT 01-OCT-1989, integrated into UniProtKB/Swiss-Prot. DT 01-OCT-1989, sequence version 1. DT 27-MAR-2024, entry version 139. DE RecName: Full=Cholesterol oxidase; DE Short=CHOD; DE EC=1.1.3.6 {ECO:0000269|PubMed:9922167}; DE AltName: Full=Cholesterol isomerase; DE EC=5.3.3.1 {ECO:0000269|PubMed:9922167}; DE Flags: Precursor; GN Name=choA; OS Streptomyces sp. (strain SA-COO). OC Bacteria; Actinomycetota; Actinomycetes; Kitasatosporales; OC Streptomycetaceae; Streptomyces. OX NCBI_TaxID=74576; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RX PubMed=2914858; DOI=10.1128/jb.171.1.596-601.1989; RA Ishizaki T., Hirayama N., Shinkawa H., Nimi O., Murooka Y.; RT "Nucleotide sequence of the gene for cholesterol oxidase from a RT Streptomyces sp."; RL J. Bacteriol. 171:596-601(1989). RN [2] RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND RP MUTAGENESIS OF GLU-398 AND HIS-484. RX PubMed=9922167; DOI=10.1021/bi982115+; RA Kass I.J., Sampson N.S.; RT "Evaluation of the role of His447 in the reaction catalyzed by cholesterol RT oxidase."; RL Biochemistry 37:17990-18000(1998). RN [3] {ECO:0007744|PDB:1B4V, ECO:0007744|PDB:1B8S, ECO:0007744|PDB:1CBO, ECO:0007744|PDB:1CC2} RP X-RAY CRYSTALLOGRAPHY (1.5 ANGSTROMS) OF WILD-TYPE AND MUTANTS GLN-398; RP ASN-484 AND GLN-484 IN COMPLEX WITH FAD, COFACTOR, AND ACTIVE SITE. RX PubMed=10194345; DOI=10.1021/bi982497j; RA Yue Q.K., Kass I.J., Sampson N.S., Vrielink A.; RT "Crystal structure determination of cholesterol oxidase from Streptomyces RT and structural characterization of key active site mutants."; RL Biochemistry 38:4277-4286(1999). CC -!- FUNCTION: Bifunctional enzyme that catalyzes the oxidation of the 3- CC beta-hydroxy group of cholesterol and the isomerization of the double CC bond of the resulting product, producing cholest-4-en-3-one. These CC reactions are part of a cholesterol degradation pathway that allows the CC bacterium to utilize cholesterol as its sole source of carbon and CC energy. {ECO:0000269|PubMed:9922167}. CC -!- CATALYTIC ACTIVITY: CC Reaction=cholesterol + O2 = cholest-5-en-3-one + H2O2; CC Xref=Rhea:RHEA:32183, ChEBI:CHEBI:15379, ChEBI:CHEBI:16113, CC ChEBI:CHEBI:16240, ChEBI:CHEBI:63906; EC=1.1.3.6; CC Evidence={ECO:0000269|PubMed:9922167}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:32184; CC Evidence={ECO:0000305|PubMed:9922167}; CC -!- CATALYTIC ACTIVITY: CC Reaction=cholest-5-en-3-one = cholest-4-en-3-one; Xref=Rhea:RHEA:32187, CC ChEBI:CHEBI:16175, ChEBI:CHEBI:63906; EC=5.3.3.1; CC Evidence={ECO:0000269|PubMed:9922167}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:32188; CC Evidence={ECO:0000305|PubMed:9922167}; CC -!- COFACTOR: CC Name=FAD; Xref=ChEBI:CHEBI:57692; CC Evidence={ECO:0000269|PubMed:10194345}; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=3 uM for cholesterol {ECO:0000269|PubMed:9922167}; CC KM=6.2 uM for cholest-5-en-3-one {ECO:0000269|PubMed:9922167}; CC Note=kcat is 44 sec(-1) for the oxidation of cholesterol. kcat is 64 CC sec(-1) for the isomerization of cholest-5-en-3-one. CC {ECO:0000269|PubMed:9922167}; CC -!- PATHWAY: Steroid metabolism; cholesterol degradation. CC {ECO:0000305|PubMed:9922167}. CC -!- SUBUNIT: Monomer. CC -!- SUBCELLULAR LOCATION: Secreted. CC -!- PTM: Predicted to be exported by the Tat system. The position of the CC signal peptide cleavage has been experimentally proven. CC -!- SIMILARITY: Belongs to the GMC oxidoreductase family. {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; M31939; AAA26719.1; -; Genomic_DNA. DR PIR; A32260; A32260. DR PDB; 1B4V; X-ray; 1.50 A; A=43-546. DR PDB; 1B8S; X-ray; 1.65 A; A=43-546. DR PDB; 1CBO; X-ray; 1.80 A; A=43-546. DR PDB; 1CC2; X-ray; 2.20 A; A=43-546. DR PDB; 1IJH; X-ray; 1.53 A; A=43-546. DR PDB; 1MXT; X-ray; 0.95 A; A=43-546. DR PDB; 1N1P; X-ray; 0.95 A; A=43-546. DR PDB; 1N4U; X-ray; 0.95 A; A=43-546. DR PDB; 1N4V; X-ray; 1.00 A; A=43-546. DR PDB; 1N4W; X-ray; 0.92 A; A=43-546. DR PDB; 2GEW; X-ray; 0.97 A; A=43-546. DR PDB; 3B3R; X-ray; 0.98 A; A=42-546. DR PDB; 3B6D; X-ray; 1.20 A; A=43-546. DR PDB; 3CNJ; X-ray; 0.95 A; A=45-543. DR PDB; 3GYI; X-ray; 1.00 A; A=43-546. DR PDB; 3GYJ; X-ray; 0.92 A; A=43-546. DR PDB; 4REK; X-ray; 0.74 A; A=46-544. DR PDB; 4U2L; X-ray; 1.34 A; A=43-546. DR PDB; 4U2S; X-ray; 1.12 A; A=43-546. DR PDB; 4U2T; X-ray; 1.22 A; A=43-546. DR PDB; 4XWR; X-ray; 1.10 A; A=43-546. DR PDB; 4XXG; X-ray; 0.85 A; A=43-546. DR PDB; 5KWF; Other; 1.50 A; A=43-546. DR PDBsum; 1B4V; -. DR PDBsum; 1B8S; -. DR PDBsum; 1CBO; -. DR PDBsum; 1CC2; -. DR PDBsum; 1IJH; -. DR PDBsum; 1MXT; -. DR PDBsum; 1N1P; -. DR PDBsum; 1N4U; -. DR PDBsum; 1N4V; -. DR PDBsum; 1N4W; -. DR PDBsum; 2GEW; -. DR PDBsum; 3B3R; -. DR PDBsum; 3B6D; -. DR PDBsum; 3CNJ; -. DR PDBsum; 3GYI; -. DR PDBsum; 3GYJ; -. DR PDBsum; 4REK; -. DR PDBsum; 4U2L; -. DR PDBsum; 4U2S; -. DR PDBsum; 4U2T; -. DR PDBsum; 4XWR; -. DR PDBsum; 4XXG; -. DR PDBsum; 5KWF; -. DR AlphaFoldDB; P12676; -. DR SMR; P12676; -. DR DrugBank; DB03147; Flavin adenine dinucleotide. DR DrugBank; DB02332; Flavin-N7 protonated-adenine dinucleotide. DR BRENDA; 1.1.3.6; 1284. DR UniPathway; UPA01058; -. DR EvolutionaryTrace; P12676; -. DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell. DR GO; GO:0016995; F:cholesterol oxidase activity; IEA:UniProtKB-EC. DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IEA:InterPro. DR GO; GO:0004769; F:steroid delta-isomerase activity; IEA:UniProtKB-EC. DR GO; GO:0006707; P:cholesterol catabolic process; IEA:UniProtKB-UniPathway. DR Gene3D; 3.50.50.60; FAD/NAD(P)-binding domain; 1. DR InterPro; IPR036188; FAD/NAD-bd_sf. DR InterPro; IPR000172; GMC_OxRdtase_N. DR InterPro; IPR007867; GMC_OxRtase_C. DR InterPro; IPR006311; TAT_signal. DR PANTHER; PTHR47470; CHOLESTEROL OXIDASE; 1. DR PANTHER; PTHR47470:SF1; FAD-DEPENDENT OXIDOREDUCTASE 2 FAD BINDING DOMAIN-CONTAINING PROTEIN; 1. DR Pfam; PF05199; GMC_oxred_C; 1. DR Pfam; PF00732; GMC_oxred_N; 1. DR SUPFAM; SSF54373; FAD-linked reductases, C-terminal domain; 1. DR SUPFAM; SSF51905; FAD/NAD(P)-binding domain; 1. DR PROSITE; PS00623; GMC_OXRED_1; 1. DR PROSITE; PS51318; TAT; 1. PE 1: Evidence at protein level; KW 3D-structure; Cholesterol metabolism; FAD; Flavoprotein; Isomerase; KW Lipid metabolism; Oxidoreductase; Secreted; Signal; Steroid metabolism; KW Sterol metabolism. FT SIGNAL 1..42 FT /note="Tat-type signal" FT CHAIN 43..546 FT /note="Cholesterol oxidase" FT /id="PRO_0000012333" FT ACT_SITE 398 FT /note="Proton acceptor" FT /evidence="ECO:0000305|PubMed:10194345" FT ACT_SITE 484 FT /note="Proton acceptor" FT /evidence="ECO:0000305|PubMed:10194345" FT BINDING 57..58 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1B4V" FT BINDING 77 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1B4V" FT BINDING 152 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1B4V" FT BINDING 156..159 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1B4V" FT BINDING 287 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1B4V" FT BINDING 483 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1IJH" FT BINDING 512 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1B4V" FT BINDING 524 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0007744|PDB:1B4V" FT MUTAGEN 398 FT /note="E->Q: Mutant does not catalyze isomerization FT (>10000-fold reduced), and oxidation activity is 30-fold FT lower than wild type." FT /evidence="ECO:0000269|PubMed:9922167" FT MUTAGEN 484 FT /note="H->D,E: Mutants do not catalyze oxidation FT (>100000-fold reduced), but do catalyze isomerization, FT 10000 times slower than wild type." FT /evidence="ECO:0000269|PubMed:9922167" FT MUTAGEN 484 FT /note="H->K: Mutant is inactive in both oxidation FT (>100000-fold reduced) and isomerization assays FT (>10000-fold reduced)." FT /evidence="ECO:0000269|PubMed:9922167" FT MUTAGEN 484 FT /note="H->N: Mutant activity is 4400-fold lower than wild FT type for oxidation, and is 30-fold lower than wild type for FT isomerization." FT /evidence="ECO:0000269|PubMed:9922167" FT MUTAGEN 484 FT /note="H->Q: Mutant activity is 120-fold lower than wild FT type for oxidation, and the same as wild type for FT isomerization." FT /evidence="ECO:0000269|PubMed:9922167" FT TURN 42..44 FT /evidence="ECO:0007829|PDB:3B3R" FT STRAND 47..53 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 57..68 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 73..79 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 89..92 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 95..97 FT /evidence="ECO:0007829|PDB:4U2T" FT HELIX 100..102 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 103..105 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 116..119 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 120..122 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 132..137 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 142..146 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 151..154 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 165..171 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 177..182 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 184..191 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 199..204 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 206..208 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 209..220 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 225..227 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 230..232 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 234..241 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 249..251 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 259..262 FT /evidence="ECO:0007829|PDB:4REK" FT TURN 265..268 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 269..275 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 278..292 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 296..305 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 311..324 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 327..340 FT /evidence="ECO:0007829|PDB:4REK" FT TURN 349..352 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 361..366 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 383..387 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 395..400 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 410..417 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 425..428 FT /evidence="ECO:0007829|PDB:4REK" FT TURN 429..432 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 433..436 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 440..443 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 444..461 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 468..473 FT /evidence="ECO:0007829|PDB:1N4W" FT STRAND 476..478 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 480..484 FT /evidence="ECO:0007829|PDB:4REK" FT TURN 491..493 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 499..501 FT /evidence="ECO:0007829|PDB:4REK" FT STRAND 505..509 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 512..514 FT /evidence="ECO:0007829|PDB:4REK" FT HELIX 524..542 FT /evidence="ECO:0007829|PDB:4REK" SQ SEQUENCE 546 AA; 58994 MW; EF22A1FE5EA68D21 CRC64; MTAQQHLSRR RMLGMAAFGA AALAGGTTIA APRAAAAAKS AADNGGYVPA VVIGTGYGAA VSALRLGEAG VQTLMLEMGQ LWNQPGPDGN IFCGMLNPDK RSSWFKNRTE APLGSFLWLD VVNRNIDPYA GVLDRVNYDQ MSVYVGRGVG GGSLVNGGMA VEPKRSYFEE ILPRVDSSEM YDRYFPRANS MLRVNHIDTK WFEDTEWYKF ARVSREQAGK AGLGTVFVPN VYDFGYMQRE AAGEVPKSAL ATEVIYGNNH GKQSLDKTYL AAALGTGKVT IQTLHQVKTI RQTKDGGYAL TVEQKDTDGK LLATKEISCR YLFLGAGSLG STELLVRARD TGTLPNLNSE VGAGWGPNGN IMTARANHMW NPTGAHQSSI PALGIDAWDN SDSSVFAEIA PMPAGLETWV SLYLAITKNP QRGTFVYDAA TDRAKLNWTR DQNAPAVNAA KALFDRINKA NGTIYRYDLF GTQLKAFADD FCYHPLGGCV LGKATDDYGR VAGYKNLYVT DGSLIPGSVG VNPFVTITAL AERNVERIIK QDVTAS //