EzCatDB: T00216
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DB codeT00216
CATH domainDomain 12.60.40.420 : Immunoglobulin-likeCatalytic domain
Domain 22.60.40.420 : Immunoglobulin-like
Domain 32.60.40.420 : Immunoglobulin-likeCatalytic domain
E.C.1.10.3.3

CATH domainRelated DB codes (homologues)
2.60.40.420 : Immunoglobulin-likeT00215,M00115,M00062,M00194

Enzyme Name
UniProtKBKEGG

P37064
Protein nameL-ascorbate oxidaseL-ascorbate oxidase
ascorbase
ascorbic acid oxidase
ascorbate oxidase
ascorbic oxidase
ascorbate dehydrogenase
L-ascorbic acid oxidase
AAO
L-ascorbate:O2 oxidoreductase
AA oxidase
SynonymsAscorbase
ASO
EC 1.10.3.3
PfamPF00394 (Cu-oxidase)
PF07731 (Cu-oxidase_2)
PF07732 (Cu-oxidase_3)
[Graphical view]

KEGG pathways
MAP codePathways
MAP00053Ascorbate and aldarate metabolism

UniProtKB:Accession NumberP37064
Entry nameASO_CUCPM
Activity2 L-ascorbate + O(2) = 2 dehydroascorbate + 2 H(2)O.
SubunitDimer.
Subcellular locationSecreted (Potential).
CofactorBinds 4 copper ions per monomer. The copper ions are bound as 3 distinct copper centers known as type 1 or blue, type 2 or normal, and type 3 or coupled binuclear.

Compound table: links to PDB-related databases & PoSSuM

CofactorsSubstratesProductsintermediates
KEGG-idC00070C00072C00007C05422C00001

CompoundCopperL-AscorbateO2DehydroascorbateH2OHydro-peroxide intermediateOxygen radical intermediate
Typeheavy metalcarbohydrate,aromatic ring (with hetero atoms other than nitrogen atoms)otherscarbohydrateH2O

ChEBI28694
30052
29073
27140
26689
15379
27956
15377


PubChem23978
54670067
977
440667
962
22247451


               
1aozA01Analogue:C2OUnboundUnboundUnbound UnboundUnbound
1aozB01Analogue:C2OUnboundUnboundUnbound UnboundUnbound
1asoA01Bound:_CUUnboundUnboundUnbound UnboundUnbound
1asoB01Bound:_CUUnboundUnboundUnbound UnboundUnbound
1aspA01Bound:_CUUnboundUnboundUnbound UnboundUnbound
1aspB01Bound:_CUUnboundUnboundUnbound UnboundUnbound
1asqA01Bound:_CUUnboundUnboundUnbound UnboundUnbound
1asqB01Bound:_CUUnboundUnboundUnbound UnboundUnbound
1aozA02UnboundUnboundUnboundUnbound UnboundUnbound
1aozB02UnboundUnboundUnboundUnbound UnboundUnbound
1asoA02UnboundUnboundUnboundUnbound UnboundUnbound
1asoB02UnboundUnboundUnboundUnbound UnboundUnbound
1aspA02UnboundUnboundUnboundUnbound UnboundUnbound
1aspB02UnboundUnboundUnboundUnbound UnboundUnbound
1asqA02UnboundUnboundUnboundUnbound UnboundUnbound
1asqB02UnboundUnboundUnboundUnbound UnboundUnbound
1aozA03Bound:_CU,Analogue;C1OUnboundUnboundUnbound UnboundUnbound
1aozB03Bound:_CU,Analogue;C1OUnboundUnboundUnbound UnboundUnbound
1asoA03Bound:3x_CUUnboundUnboundUnbound UnboundUnbound
1asoB03Bound:3x_CUUnboundUnboundUnbound UnboundUnbound
1aspA03Bound:3x_CUUnboundUnboundUnbound Bound:PEOUnbound
1aspB03Bound:3x_CUUnboundUnboundUnbound Bound:PEOUnbound
1asqA03Bound:3x_CUUnboundUnboundUnbound Analogue:AZI 2Unbound
1asqB03Bound:3x_CUUnboundUnboundUnbound Analogue:AZI 2Unbound

Active-site residues
resource
literature [7], [8]
pdbCatalytic residuesCofactor-binding residues
          
1aozA01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1aozB01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1asoA01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1asoB01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1aspA01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1aspB01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1asqA01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1asqB01 
HIS 106(Copper-3a);HIS 62;HIS 104(Copper-3b);HIS 60(Copper-2)
1aozA02 
 
1aozB02 
 
1asoA02 
 
1asoB02 
 
1aspA02 
 
1aspB02 
 
1asqA02 
 
1asqB02 
 
1aozA03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)
1aozB03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)
1asoA03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)
1asoB03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)
1aspA03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)
1aspB03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)
1asqA03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)
1asqB03HIS 506;CYS 507;HIS 508
HIS 445;CYS 507;HIS 512;MET 517(Copper-1);HIS 450;HIS 506(Copper-3a);HIS 508(Copper-3b);HIS 448(Copper-2)

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[7]Fig.19, p.199-203
[8]Fig.13, p.1012-1013

references
[1]
PubMed ID510553
JournalFEBS Lett
Year1979
Volume107
Pages407-8
AuthorsLadenstein R, Marchesini A, Palmieri S
TitlePreliminary crystallographic study of ascorbic acid oxidase from green zucchini squash.
[2]
PubMed ID6620382
JournalJ Mol Biol
Year1983
Volume169
Pages351-2
AuthorsBolognesi M, Gatti G, Coda A, Avigliano L, Marcozzi G, Finazzi-Agro A
TitleA new crystalline modification of the copper enzyme ascorbate oxidase.
[3]
PubMed ID3406029
JournalProg Clin Biol Res
Year1988
Volume274
Pages285-8
AuthorsMesserschmidt A, Rossi A, Ladenstein R, Huber R, Bolognesi M, Marchesini A, Petruzelli R, Finazzi-Agro A
TitlePreliminary X-ray crystal structure and partial cDNA-sequence of ascorbate oxidase from zucchini.
[4]
CommentsX-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS)
Medline ID89236417
PubMed ID2716059
JournalJ Mol Biol
Year1989
Volume206
Pages513-29
AuthorsMesserschmidt A, Rossi A, Ladenstein R, Huber R, Bolognesi M, Gatti G, Marchesini A, Petruzzelli R, Finazzi-Agro A
TitleX-ray crystal structure of the blue oxidase ascorbate oxidase from zucchini. Analysis of the polypeptide fold and a model of the copper sites and ligands.
Related UniProtKBP37064
[5]
PubMed ID2373076
JournalEur J Biochem
Year1990
Volume190
Pages491-5
AuthorsSavini I, D'Alessio S, Giartosio A, Morpurgo L, Avigliano L
TitleThe role of copper in the stability of ascorbate oxidase towards denaturing agents.
[6]
PubMed ID1330552
JournalEur J Biochem
Year1992
Volume209
Pages597-602
AuthorsMesserschmidt A, Steigemann W, Huber R, Lang G, Kroneck PM
TitleX-ray crystallographic characterisation of type-2-depleted ascorbate oxidase from zucchini.
[7]
CommentsX-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS)
Medline ID92194315
PubMed ID1548698
JournalJ Mol Biol
Year1992
Volume224
Pages179-205
AuthorsMesserschmidt A, Ladenstein R, Huber R, Bolognesi M, Avigliano L, Petruzzelli R, Rossi A, Finazzi-Agro A
TitleRefined crystal structure of ascorbate oxidase at 1.9 A resolution.
Related PDB1aoz
Related UniProtKBP37064
[8]
CommentsX-ray crystallography
PubMed ID8478945
JournalJ Mol Biol
Year1993
Volume230
Pages997-1014
AuthorsMesserschmidt A, Luecke H, Huber R
TitleX-ray structures and mechanistic implications of three functional derivatives of ascorbate oxidase from zucchini. Reduced, peroxide and azide forms.
Related PDB1aso,1asp,1asq
[9]
PubMed ID7748896
JournalBiochim Biophys Acta
Year1995
Volume1248
Pages143-8
AuthorsSakurai T, Takahashi J
TitleEPR spectra of type 3 copper centers in Rhus vernicifera laccase and Cucumis sativus ascorbate oxidase.
[10]
PubMed ID9125505
JournalBiochemistry
Year1997
Volume36
Pages4852-9
AuthorsGaspard S, Monzani E, Casella L, Gullotti M, Maritano S, Marchesini A
TitleInhibition of ascorbate oxidase by phenolic compounds. Enzymatic and spectroscopic studies.
[11]
PubMed ID9283082
JournalBiochemistry
Year1997
Volume36
Pages10917-22
AuthorsMei G, Di Venere A, Buganza M, Vecchini P, Rosato N, Finazzi-Agro' A
TitleRole of quaternary structure in the stability of dimeric proteins: the case of ascorbate oxidase.
[12]
PubMed ID9048895
JournalBiochim Biophys Acta
Year1997
Volume1337
Pages191-7
AuthorsReinhammar B, Aasa R, Vanngard T, Maritano S, Marchesini A
TitleThe type 2 copper of ascorbate oxidase.
[13]
PubMed ID9602107
JournalBiochim Biophys Acta
Year1998
Volume1384
Pages160-70
AuthorsHuang HW, Sakurai T, Monjushiro H, Takeda S
TitleMagnetic studies of the trinuclear center in laccase and ascorbate oxidase approached by EPR spectroscopy and magnetic susceptibility measurements.
[14]
PubMed ID9781686
JournalFEBS Lett
Year1998
Volume436
Pages239-42
AuthorsFarver O, Eady RR, Abraham ZH, Pecht I
TitleThe intramolecular electron transfer between copper sites of nitrite reductase: a comparison with ascorbate oxidase.
[15]
PubMed ID10813026
JournalAppl Biochem Biotechnol
Year1999
Volume82
Pages227-41
AuthorsSavini I, Santucci R, Di Venere A, Rosato N, Strukul G, Pinna F, Avigliano L
TitleCatalytic and spectroscopic properties of cytochrome-c, horseradish peroxidase, and ascorbate oxidase embedded in a sol-gel silica matrix as a function of gelation time.
[16]
PubMed ID10583375
JournalEur J Biochem
Year1999
Volume266
Pages820-30
AuthorsGromov I, Marchesini A, Farver O, Pecht I, Goldfarb D
TitleAzide binding to the trinuclear copper center in laccase and ascorbate oxidase.
[17]
PubMed ID10860754
JournalJ Mol Biol
Year2000
Volume299
Pages487-98
AuthorsFleming PJ, Richards FM
TitleProtein packing: dependence on protein size, secondary structure and amino acid composition.
[18]
PubMed ID11983419
JournalBiochim Biophys Acta
Year2002
Volume1596
Pages36-46
AuthorsSugino M, Kajita S, Banno K, Shirai T, Yamane T, Kato M, Kobayashi T, Tsukagoshi N
TitleUpward shift of the pH optimum of Acremonium ascorbate oxidase.
[19]
PubMed ID15006640
JournalInt J Biochem Cell Biol
Year2004
Volume36
Pages881-92
AuthorsSantagostini L, Gullotti M, De Gioia L, Fantucci P, Franzini E, Marchesini A, Monzani E, Casella L
TitleProbing the location of the substrate binding site of ascorbate oxidase near type 1 copper: an investigation through spectroscopic, inhibition and docking studies.

comments
Coppers can be classified into three types (see literature [7], [8] & laccase;T00215 in EzCatDB):
Type I copper (blue copper); Copper-1 in this enzyme
Type II copper (non-blue copper); Copper-2
Type III copper (EPR-non-detectable copper); Copper-3a & copper-3b in this enzyme.
The copper-2, -3a and -3b form a trinuclear copper site, to which an oxygen is bound, whereas the cooper-1 is a monocuclear copper site, to which an organic substrate is bound (see laccase;T00215).
This enzyme catalyzes the following reactions:
(A) Oxidation (dehydrogenation) of organic substrate at mononuclear coper site:
(B) Electron transfer from mononuclear copper site to trinuclear copper site:
(C) O2 reduction to produce H2O at trinuclear copper site:

createdupdated
2005-04-222009-02-26
Copyright: Nozomi Nagano, JST & CBRC-AIST
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Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2005 - March 2006)
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Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2012 - March 2013)
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