DB code: T00216

CATH domain 2.60.40.420 : Immunoglobulin-like Catalytic domain
2.60.40.420 : Immunoglobulin-like
2.60.40.420 : Immunoglobulin-like Catalytic domain
E.C. 1.10.3.3
CSA
M-CSA
MACiE

CATH domain Related DB codes (homologues)
2.60.40.420 : Immunoglobulin-like T00215 M00115 M00062 M00194

Uniprot Enzyme Name
UniprotKB Protein name Synonyms Pfam
P37064 L-ascorbate oxidase
Ascorbase
ASO
EC 1.10.3.3
PF00394 (Cu-oxidase)
PF07731 (Cu-oxidase_2)
PF07732 (Cu-oxidase_3)
[Graphical View]

KEGG enzyme name
L-ascorbate oxidase
ascorbase
ascorbic acid oxidase
ascorbate oxidase
ascorbic oxidase
ascorbate dehydrogenase
L-ascorbic acid oxidase
AAO
L-ascorbate:O2 oxidoreductase
AA oxidase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P37064 ASO_CUCPM 2 L-ascorbate + O(2) = 2 dehydroascorbate + 2 H(2)O. Dimer. Secreted (Potential). Binds 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.

KEGG Pathways
Map code Pathways E.C.
MAP00053 Ascorbate and aldarate metabolism

Compound table
Cofactors Substrates Products Intermediates
KEGG-id C00070 C00072 C00007 C05422 C00001
E.C.
Compound Copper L-Ascorbate O2 Dehydroascorbate H2O Hydro-peroxide intermediate Oxygen radical intermediate
Type heavy metal carbohydrate,aromatic ring (with hetero atoms other than nitrogen atoms) others carbohydrate H2O
ChEBI 28694
 30052
 28694
 30052
 		29073
 29073
 		15379
 26689
 27140
 15379
 26689
 27140
 		27956
 27956
 		15377
 15377
PubChem 23978
 23978
 		54670067
 54670067
 		977
 977
 		440667
 440667
 		22247451
 962
 22247451
 962
1aozA01 Analogue:C2O Unbound Unbound Unbound Unbound Unbound
1aozB01 Analogue:C2O Unbound Unbound Unbound Unbound Unbound
1asoA01 Bound:_CU Unbound Unbound Unbound Unbound Unbound
1asoB01 Bound:_CU Unbound Unbound Unbound Unbound Unbound
1aspA01 Bound:_CU Unbound Unbound Unbound Unbound Unbound
1aspB01 Bound:_CU Unbound Unbound Unbound Unbound Unbound
1asqA01 Bound:_CU Unbound Unbound Unbound Unbound Unbound
1asqB01 Bound:_CU Unbound Unbound Unbound Unbound Unbound
1aozA02 Unbound Unbound Unbound Unbound Unbound Unbound
1aozB02 Unbound Unbound Unbound Unbound Unbound Unbound
1asoA02 Unbound Unbound Unbound Unbound Unbound Unbound
1asoB02 Unbound Unbound Unbound Unbound Unbound Unbound
1aspA02 Unbound Unbound Unbound Unbound Unbound Unbound
1aspB02 Unbound Unbound Unbound Unbound Unbound Unbound
1asqA02 Unbound Unbound Unbound Unbound Unbound Unbound
1asqB02 Unbound Unbound Unbound Unbound Unbound Unbound
1aozA03 Bound:_CU,Analogue;C1O Unbound Unbound Unbound Unbound Unbound
1aozB03 Bound:_CU,Analogue;C1O Unbound Unbound Unbound Unbound Unbound
1asoA03 Bound:3x_CU Unbound Unbound Unbound Unbound Unbound
1asoB03 Bound:3x_CU Unbound Unbound Unbound Unbound Unbound
1aspA03 Bound:3x_CU Unbound Unbound Unbound Bound:PEO Unbound
1aspB03 Bound:3x_CU Unbound Unbound Unbound Bound:PEO Unbound
1asqA03 Bound:3x_CU Unbound Unbound Unbound Analogue:AZI_2 Unbound
1asqB03 Bound:3x_CU Unbound Unbound Unbound Analogue:AZI_2 Unbound

Reference for Active-site residues
resource references E.C.
literature [7], [8]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
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
1aozA03 HIS 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)
1aozB03 HIS 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)
1asoA03 HIS 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)
1asoB03 HIS 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)
1aspA03 HIS 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)
1aspB03 HIS 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)
1asqA03 HIS 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)
1asqB03 HIS 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
References Sections No. of steps in catalysis
[7]
Fig.19, p.199-203
[8]
Fig.13, p.1012-1013

References
[1]
Resource
Comments
Medline ID
PubMed ID 510553
Journal FEBS Lett
Year 1979
Volume 107
Pages 407-8
Authors Ladenstein R, Marchesini A, Palmieri S
Title Preliminary crystallographic study of ascorbic acid oxidase from green zucchini squash.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 6620382
Journal J Mol Biol
Year 1983
Volume 169
Pages 351-2
Authors Bolognesi M, Gatti G, Coda A, Avigliano L, Marcozzi G, Finazzi-Agro A
Title A new crystalline modification of the copper enzyme ascorbate oxidase.
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 3406029
Journal Prog Clin Biol Res
Year 1988
Volume 274
Pages 285-8
Authors Messerschmidt A, Rossi A, Ladenstein R, Huber R, Bolognesi M, Marchesini A, Petruzelli R, Finazzi-Agro A
Title Preliminary X-ray crystal structure and partial cDNA-sequence of ascorbate oxidase from zucchini.
Related PDB
Related UniProtKB
[4]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS)
Medline ID 89236417
PubMed ID 2716059
Journal J Mol Biol
Year 1989
Volume 206
Pages 513-29
Authors Messerschmidt A, Rossi A, Ladenstein R, Huber R, Bolognesi M, Gatti G, Marchesini A, Petruzzelli R, Finazzi-Agro A
Title X-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 PDB
Related UniProtKB P37064
[5]
Resource
Comments
Medline ID
PubMed ID 2373076
Journal Eur J Biochem
Year 1990
Volume 190
Pages 491-5
Authors Savini I, D'Alessio S, Giartosio A, Morpurgo L, Avigliano L
Title The role of copper in the stability of ascorbate oxidase towards denaturing agents.
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 1330552
Journal Eur J Biochem
Year 1992
Volume 209
Pages 597-602
Authors Messerschmidt A, Steigemann W, Huber R, Lang G, Kroneck PM
Title X-ray crystallographic characterisation of type-2-depleted ascorbate oxidase from zucchini.
Related PDB
Related UniProtKB
[7]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS)
Medline ID 92194315
PubMed ID 1548698
Journal J Mol Biol
Year 1992
Volume 224
Pages 179-205
Authors Messerschmidt A, Ladenstein R, Huber R, Bolognesi M, Avigliano L, Petruzzelli R, Rossi A, Finazzi-Agro A
Title Refined crystal structure of ascorbate oxidase at 1.9 A resolution.
Related PDB 1aoz
Related UniProtKB P37064
[8]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 8478945
Journal J Mol Biol
Year 1993
Volume 230
Pages 997-1014
Authors Messerschmidt A, Luecke H, Huber R
Title X-ray structures and mechanistic implications of three functional derivatives of ascorbate oxidase from zucchini. Reduced, peroxide and azide forms.
Related PDB 1aso 1asp 1asq
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 7748896
Journal Biochim Biophys Acta
Year 1995
Volume 1248
Pages 143-8
Authors Sakurai T, Takahashi J
Title EPR spectra of type 3 copper centers in Rhus vernicifera laccase and Cucumis sativus ascorbate oxidase.
Related PDB
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 9125505
Journal Biochemistry
Year 1997
Volume 36
Pages 4852-9
Authors Gaspard S, Monzani E, Casella L, Gullotti M, Maritano S, Marchesini A
Title Inhibition of ascorbate oxidase by phenolic compounds. Enzymatic and spectroscopic studies.
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 9283082
Journal Biochemistry
Year 1997
Volume 36
Pages 10917-22
Authors Mei G, Di Venere A, Buganza M, Vecchini P, Rosato N, Finazzi-Agro' A
Title Role of quaternary structure in the stability of dimeric proteins: the case of ascorbate oxidase.
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 9048895
Journal Biochim Biophys Acta
Year 1997
Volume 1337
Pages 191-7
Authors Reinhammar B, Aasa R, Vanngard T, Maritano S, Marchesini A
Title The type 2 copper of ascorbate oxidase.
Related PDB
Related UniProtKB
[13]
Resource
Comments
Medline ID
PubMed ID 9602107
Journal Biochim Biophys Acta
Year 1998
Volume 1384
Pages 160-70
Authors Huang HW, Sakurai T, Monjushiro H, Takeda S
Title Magnetic studies of the trinuclear center in laccase and ascorbate oxidase approached by EPR spectroscopy and magnetic susceptibility measurements.
Related PDB
Related UniProtKB
[14]
Resource
Comments
Medline ID
PubMed ID 9781686
Journal FEBS Lett
Year 1998
Volume 436
Pages 239-42
Authors Farver O, Eady RR, Abraham ZH, Pecht I
Title The intramolecular electron transfer between copper sites of nitrite reductase: a comparison with ascorbate oxidase.
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 10813026
Journal Appl Biochem Biotechnol
Year 1999
Volume 82
Pages 227-41
Authors Savini I, Santucci R, Di Venere A, Rosato N, Strukul G, Pinna F, Avigliano L
Title Catalytic and spectroscopic properties of cytochrome-c, horseradish peroxidase, and ascorbate oxidase embedded in a sol-gel silica matrix as a function of gelation time.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 10583375
Journal Eur J Biochem
Year 1999
Volume 266
Pages 820-30
Authors Gromov I, Marchesini A, Farver O, Pecht I, Goldfarb D
Title Azide binding to the trinuclear copper center in laccase and ascorbate oxidase.
Related PDB
Related UniProtKB
[17]
Resource
Comments
Medline ID
PubMed ID 10860754
Journal J Mol Biol
Year 2000
Volume 299
Pages 487-98
Authors Fleming PJ, Richards FM
Title Protein packing: dependence on protein size, secondary structure and amino acid composition.
Related PDB
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 11983419
Journal Biochim Biophys Acta
Year 2002
Volume 1596
Pages 36-46
Authors Sugino M, Kajita S, Banno K, Shirai T, Yamane T, Kato M, Kobayashi T, Tsukagoshi N
Title Upward shift of the pH optimum of Acremonium ascorbate oxidase.
Related PDB
Related UniProtKB
[19]
Resource
Comments
Medline ID
PubMed ID 15006640
Journal Int J Biochem Cell Biol
Year 2004
Volume 36
Pages 881-92
Authors Santagostini L, Gullotti M, De Gioia L, Fantucci P, Franzini E, Marchesini A, Monzani E, Casella L
Title Probing the location of the substrate binding site of ascorbate oxidase near type 1 copper: an investigation through spectroscopic, inhibition and docking studies.
Related PDB
Related UniProtKB

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:

Created Updated
2005-04-22 2009-02-26