CHOD_STRS0
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 03-AUG-2022, entry version 134.
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; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC 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=3.0 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}.
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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; -; 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 Pfam; PF05199; GMC_oxred_C; 1.
DR Pfam; PF00732; GMC_oxred_N; 1.
DR SUPFAM; SSF51905; SSF51905; 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 (>100000-
FT fold reduced), but do catalyze isomerization, 10000 times
FT slower than wild type."
FT /evidence="ECO:0000269|PubMed:9922167"
FT MUTAGEN 484
FT /note="H->K: Mutant is inactive in both oxidation (>100000-
FT fold reduced) and isomerization assays (>10000-fold
FT 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
