EzCatDB: D00015

DB codeD00015
CATH domainDomain 13.50.50.60Catalytic domain
Domain 23.30.410.10Catalytic domain
E.C.1.1.3.6
CSA1coy

CATH domainRelated DB codes (homologues)
3.50.50.60M00163,D00041,D00042,D00045,D00064,D00071,T00004,T00015,T00017,T00025,T00211,T00213,T00233,T00242

Enzyme Name
Swiss-protKEGG

P22637P12676
Protein nameCholesterol oxidaseCholesterol oxidasecholesterol oxidase
cholesterol- O2 oxidoreductase
3beta-hydroxy steroid oxidoreductase
3beta-hydroxysteroid:oxygen oxidoreductase
SynonymsCHOD
EC 1.1.3.6
CHOD
EC 1.1.3.6

KEGG pathways
MAP codePathways
MAP00120Bile acid biosynthesis

Swiss-prot:Accession NumberP22637P12676
Entry nameCHOD_BRESTCHOD_STRS0
ActivityCholesterol + O(2) = cholest-4-en-3-one + H(2)O(2).Cholesterol + O(2) = cholest-4-en-3-one + H(2)O(2).
Subunit
Monomer.
Subcellular locationSecreted.Secreted.
CofactorFAD.FAD.


CofactorsSubstratesProductsintermediates
KEGG-idC00016C00187C00007C00599C00027
CompoundFADCholesterolO2Cholest-4-en-3-oneH2O25-cholesten-3-one
Typeamide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carbohydrate,nucleotidelipid,steroidotherscarbohydrate,lipid,steroidothers
1b4vA01Bound:FADUnboundUnboundUnboundUnboundUnbound
1b8sA01Bound:FADUnboundUnboundUnboundUnboundUnbound
1cboA01Bound:FADUnboundUnboundUnboundUnboundUnbound
1cc2A01Bound:FADUnboundUnboundUnboundUnboundUnbound
1coxA01UnboundUnboundUnboundUnboundUnboundUnbound
1coyA01Bound:FADAnalogue:ANDUnboundUnboundUnboundUnbound
1ijhA01Bound:FADUnboundUnboundUnboundUnboundUnbound
1mxtA01Analogue:FAEUnboundBound:OXYUnboundUnboundUnbound
1n1pA01Bound:FADUnboundUnboundUnboundUnboundUnbound
1n4uA01Analogue:FAEUnboundBound:OXYUnboundUnboundUnbound
1n4vA01Bound:FADUnboundUnboundUnboundUnboundUnbound
1n4wA01Bound:FADUnboundUnboundUnboundUnboundUnbound
3coxA01Bound:FADUnboundUnboundUnboundUnboundUnbound
1b4vA02UnboundUnboundUnboundUnboundUnboundUnbound
1b8sA02UnboundUnboundUnboundUnboundUnboundUnbound
1cboA02UnboundUnboundUnboundUnboundUnboundUnbound
1cc2A02UnboundUnboundUnboundUnboundUnboundUnbound
1coxA02UnboundUnboundUnboundUnboundUnboundUnbound
1coyA02UnboundUnboundUnboundUnboundUnboundUnbound
1ijhA02UnboundUnboundUnboundUnboundUnboundUnbound
1mxtA02UnboundUnboundUnboundUnboundUnboundUnbound
1n1pA02UnboundUnboundUnboundUnboundUnboundUnbound
1n4uA02UnboundUnboundUnboundUnboundUnboundUnbound
1n4vA02UnboundUnboundUnboundUnboundUnboundUnbound
1n4wA02UnboundUnboundUnboundUnboundUnboundUnbound
3coxA02UnboundUnboundUnboundUnboundUnboundUnbound

Active-site residues
resource
PDB;1b4v, 1b8s, 1cbo, 1cc2 & Swiss-prot;P12676,P22637 & literature [19]
pdbCatalytic residuescomment
1b4vA01HIS 447;ASN 485

1b8sA01HIS 447;ASN 485

1cboA01       ;ASN 485
mutant H447N
1cc2A01       ;ASN 485
mutant H447Q
1coxA01HIS 447;ASN 485

1coyA01HIS 447;ASN 485

1ijhA01HIS 447;       
mutant N485L
1mxtA01HIS 447;ASN 485

1n1pA01HIS 447;ASN 485

1n4uA01HIS 447;ASN 485

1n4vA01HIS 447;ASN 485

1n4wA01HIS 447;ASN 485

3coxA01HIS 447;ASN 485

1b4vA02GLU 361

1b8sA02       
mutant E361Q
1cboA02GLU 361

1cc2A02GLU 361

1coxA02GLU 361

1coyA02GLU 361

1ijhA02GLU 361

1mxtA02GLU 361

1n1pA02GLU 361

1n4uA02GLU 361

1n4vA02GLU 361

1n4wA02GLU 361

3coxA02GLU 361


References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[2]Fig.1, p.551-552
[3]Fig.1, Fig.8, Fig.9, p.11512-11514
[8]p.17995-17999
[11]Fig.5, Fig.6, p.1079-1080
[12]Scheme 1, p.622
[13]p.4282-4285
[14]p.149-150
[16]Fig.1
[17]Fig.1, Fig.4
[18]FIG. 1, p.841-842
[19]p.13785-13785
[20]p.30438-30440
[23]Fig.1, p.713-715
[25]Scheme 2, p.1640-1647

references
[1]
PubMed ID6528973
JournalAnal Biochem
Year1984
Volume142
Pages347-50
AuthorsOmodeo Sale F, Marchesini S, Fishman PH, Berra B
TitleA sensitive enzymatic assay for determination of cholesterol in lipid extracts.
[2]
CommentsX-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS)
Medline ID91269328
PubMed ID2051487
JournalJ Mol Biol
Year1991
Volume219
Pages533-54
AuthorsVrielink A, Lloyd LF, Blow DM
TitleCrystal structure of cholesterol oxidase from Brevibacterium sterolicum refined at 1.8 A resolution.
Related PDB1cox
Related Swiss-protP22637
[3]
CommentsX-ray crystallography
PubMed ID8218217
JournalBiochemistry
Year1993
Volume32
Pages11507-15
AuthorsLi J, Vrielink A, Brick P, Blow DM
TitleCrystal structure of cholesterol oxidase complexed with a steroid substrate: implications for flavin adenine dinucleotide dependent alcohol oxidases.
Related PDB3cox,1coy
[4]
PubMed ID8459768
JournalMol Microbiol
Year1993
Volume7
Pages419-28
AuthorsMolnar I, Hayashi N, Choi KP, Yamamoto H, Yamashita M, Murooka Y
TitleBacterial cholesterol oxidases are able to act as flavoprotein-linked ketosteroid monooxygenases that catalyse the hydroxylation of cholesterol to 4-cholesten-6-ol-3-one.
[5]
PubMed ID8812988
JournalJ Struct Biol
Year1996
Volume116
Pages317-9
AuthorsCroteau N, Vrielink A
TitleCrystallization and preliminary X-ray analysis of cholesterol oxidase from Brevibacterium sterolicum containing covalently bound FAD.
[6]
PubMed ID9001407
JournalFEBS Lett
Year1997
Volume400
Pages247-51
AuthorsMedina M, Vrielink A, Cammack R
TitleElectron spin echo envelope modulation studies of the semiquinone anion radical of cholesterol oxidase from Brevibacterium sterolicum.
[7]
PubMed ID9153088
JournalProtein Eng
Year1997
Volume10
Pages231-5
AuthorsNishiya Y, Harada N, Teshima SI, Yamashita M, Fujii I, Hirayama N, Murooka Y
TitleImprovement of thermal stability of Streptomyces cholesterol oxidase by random mutagenesis and a structural interpretation.
[8]
PubMed ID9922167
JournalBiochemistry
Year1998
Volume37
Pages17990-8000
AuthorsKass IJ, Sampson NS
TitleEvaluation of the role of His447 in the reaction catalyzed by cholesterol oxidase.
[9]
PubMed ID9548964
JournalBiochemistry
Year1998
Volume37
Pages5770-8
AuthorsSampson NS, Kass IJ, Ghoshroy KB
TitleAssessment of the role of an omega loop of cholesterol oxidase: a truncated loop mutant has altered substrate specificity.
[10]
PubMed ID9749912
JournalProtein Eng
Year1998
Volume11
Pages609-11
AuthorsNishiya Y, Yamashita M, Murooka Y, Fujii I, Hirayama N
TitleEffect of non-ionic detergents on apparent enzyme mechanism: V121A mutant of Streptomyces cholesterol oxidase endowed with enhanced sensitivity towards detergents.
[11]
PubMed ID9876929
JournalProtein Eng
Year1998
Volume11
Pages1075-81
AuthorsYamashita M, Toyama M, Ono H, Fujii I, Hirayama N, Murooka Y
TitleSeparation of the two reactions, oxidation and isomerization, catalyzed by Streptomyces cholesterol oxidase.
[12]
PubMed ID10417325
JournalBiochem J
Year1999
Volume341
Pages621-7
AuthorsDoukyu N, Aono R
TitleTwo moles of O2 consumption and one mole of H2O2 formation during cholesterol peroxidation with cholesterol oxidase from Pseudomonas sp. strain ST-200.
[13]
CommentsX-RAY CRYSTALLOGRAPHY (1.5 ANGSTROMS), AND MUTAGENESIS
Medline ID99211873
PubMed ID10194345
JournalBiochemistry
Year1999
Volume38
Pages4277-86
AuthorsYue QK, Kass IJ, Sampson NS, Vrielink A
TitleCrystal structure determination of cholesterol oxidase from Streptomyces and structural characterization of key active site mutants.
Related PDB1b4v,1b8s,1cbo,1cc2
Related Swiss-protP12676
[14]
PubMed ID10447682
JournalEur J Biochem
Year1999
Volume264
Pages140-51
AuthorsPollegioni L, Wels G, Pilone MS, Ghisla S
TitleKinetic mechanisms of cholesterol oxidase from Streptomyces hygroscopicus and Brevibacterium sterolicum.
[15]
PubMed ID10949695
JournalAppl Biochem Biotechnol
Year2000
Volume87
Pages141-51
AuthorsYotova LK, Ivanov IP
TitleKinetic studies and analytical application of cholesterol oxidase and peroxidase immobilized to synthetic polymer.
[16]
PubMed ID10822008
JournalJ Steroid Biochem Mol Biol
Year2000
Volume72
Pages169-95
AuthorsMacLachlan J, Wotherspoon AT, Ansell RO, Brooks CJ
TitleCholesterol oxidase: sources, physical properties and analytical applications.
[17]
PubMed ID10694883
JournalTrends Biochem Sci
Year2000
Volume25
Pages126-32
AuthorsFraaije MW, Mattevi A
TitleFlavoenzymes: diverse catalysts with recurrent features.
[18]
PubMed ID11761331
JournalAntioxid Redox Signal
Year2001
Volume3
Pages839-46
AuthorsSampson NS
TitleDissection of a flavoenzyme active site: the reaction catalyzed by cholesterol oxidase.
[19]
CommentsX-ray crystallography
PubMed ID11705367
JournalBiochemistry
Year2001
Volume40
Pages13779-87
AuthorsYin Y, Sampson NS, Vrielink A, Lario PI
TitleThe presence of a hydrogen bond between asparagine 485 and the pi system of FAD modulates the redox potential in the reaction catalyzed by cholesterol oxidase.
Related PDB1ijh
[20]
PubMed ID11397813
JournalJ Biol Chem
Year2001
Volume276
Pages30435-41
AuthorsCoulombe R, Yue KQ, Ghisla S, Vrielink A
TitleOxygen access to the active site of cholesterol oxidase through a narrow channel is gated by an Arg-Glu pair.
[21]
PubMed ID11359791
JournalJ Biol Chem
Year2001
Volume276
Pages18024-30
AuthorsMotteran L, Pilone MS, Molla G, Ghisla S, Pollegioni L
TitleCholesterol oxidase from Brevibacterium sterolicum. The relationship between covalent flavinylation and redox properties.
[22]
PubMed ID12051668
JournalArch Biochem Biophys
Year2002
Volume402
Pages235-42
AuthorsYin Y, Liu P, Anderson RG, Sampson NS
TitleConstruction of a catalytically inactive cholesterol oxidase mutant: investigation of the interplay between active site-residues glutamate 361 and histidine 447.
[23]
PubMed ID12974654
JournalAcc Chem Res
Year2003
Volume36
Pages713-22
AuthorsSampson NS, Vrielink A
TitleCholesterol oxidases: a study of nature's approach to protein design.
[24]
CommentsX-ray crystallography
PubMed ID14558826
JournalJ Am Chem Soc
Year2003
Volume125
Pages12787-94
AuthorsLario PI, Vrielink A
TitleAtomic resolution density maps reveal secondary structure dependent differences in electronic distribution.
Related PDB1n1p
[25]
CommentsX-ray crystallography
PubMed ID12595270
JournalJ Mol Biol
Year2003
Volume326
Pages1635-50
AuthorsLario PI, Sampson N, Vrielink A
TitleSub-atomic resolution crystal structure of cholesterol oxidase: what atomic resolution crystallography reveals about enzyme mechanism and the role of the FAD cofactor in redox activity.
Related PDB1mxt

comments
This enzyme catalyzes three subsequent reactions;
(A) Hydride transfer from FADH2 to O2, giving FAD and H2O2.
(B) Hydride transfer from cholesterol (or 5-cholestan-3-ol) to FAD, giving 5-chelesten-3-one and FADH2.
(C) Isomerization (or shift of double-bonde) of 5-cholesten-3-one, producing 4-cholesten-3-one (shift from C4-C5=C6 to C4=C5-C6).

createdupdated
2004-03-242009-03-16


Copyright: Nozomi Nagano, JST & CBRC-AIST
Funded by PRESTO/Japan Science and Technology Corporation (JST) (December 2001 - November 2004)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
Funded by Grant-in-Aid for Scientific Research (B)/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2008)
Funded by BIRD/Japan Science and Technology Corporation (JST) (September 2005 - September 2010)
Funded by BIRD/Japan Science and Technology Corporation (JST) (October 2007 - September 2010)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2011 - March 2012)

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