EzCatDB: D00060
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DB codeD00060
CATH domainDomain 11.10.520.10 : Peroxidase; domain 1Catalytic domain
Domain 21.10.420.10 : Peroxidase; domain 2Catalytic domain
E.C.1.11.1.11
CSA1apx

CATH domainRelated DB codes (homologues)
1.10.420.10 : Peroxidase; domain 2D00061,D00062
1.10.520.10 : Peroxidase; domain 1D00061,D00062

Enzyme Name
UniProtKBKEGG

P48534Q43758
Protein nameL-ascorbate peroxidase, cytosolic
L-ascorbate peroxidase
L-ascorbic acid peroxidase
L-ascorbic acid-specific peroxidase
ascorbate peroxidase
ascorbic acid peroxidase
SynonymsAP
EC 1.11.1.11
PsAPx01
Ascorbate peroxidase
EC 1.11.1.11
Cytosolic ascorbate peroxidase 1
EC 1.11.1.11
RefSeq
NP_001237785.1 (Protein)
NM_001250856.1 (DNA/RNA sequence)
PfamPF00141 (peroxidase)
[Graphical view]
PF00141 (peroxidase)
[Graphical view]

KEGG pathways
MAP codePathways
MAP00053Ascorbate and aldarate metabolism
MAP00480Glutathione metabolism

UniProtKB:Accession NumberP48534Q43758
Entry nameAPX1_PEAQ43758_SOYBN
ActivityL-ascorbate + H(2)O(2) = dehydroascorbate + 2 H(2)O.
Subunit

Subcellular locationCytoplasm.
CofactorBinds 1 heme B (iron-protoporphyrin IX) group per subunit.,Binds 1 potassium or calcium ion per subunit.

Compound table: links to PDB-related databases & PoSSuM

CofactorsSubstratesProducts
KEGG-idC00032C00076C00238C00072C00027C05422C00001
CompoundHemeCalciumPotassiumL-AscorbateH2O2DehydroascorbateH2O
Typearomatic ring (with nitrogen atoms),carboxyl group,heavy metaldivalent metal (Ca2+, Mg2+)univalent metal (Na+, K+)carbohydrate,aromatic ring (with hetero atoms other than nitrogen atoms)otherscarbohydrateH2O
ChEBI17627
26355
29108
29103
29073
16240
27956
15377
PubChem
271
813
54670067
784
22326046
440667
962
22247451
               
1apxA01UnboundUnboundUnboundUnboundUnboundUnbound 
1apxB01UnboundUnboundUnboundUnboundUnboundUnbound 
1apxC01UnboundUnboundUnboundUnboundUnboundUnbound 
1apxD01UnboundUnboundUnboundUnboundUnboundUnbound 
1oafA01UnboundUnboundUnboundBound:ASCUnboundUnbound 
1oagA01UnboundUnboundUnboundUnboundUnboundUnbound 
1v0hX01UnboundUnboundUnboundAnalogue:SHAUnboundUnbound 
1apxA02Bound:HEMUnboundBound:__KUnboundUnboundUnbound 
1apxB02Bound:HEMUnboundBound:__KUnboundUnboundUnbound 
1apxC02Bound:HEMUnboundBound:__KUnboundUnboundUnbound 
1apxD02Bound:HEMUnboundBound:__KUnboundUnboundUnbound 
1oafA02Bound:HEMUnboundAnalogue:_NAUnboundUnboundUnbound 
1oagA02Bound:HEMUnboundUnboundUnboundUnboundUnbound 
1v0hX02Bound:HEMUnboundAnalogue:_NAUnboundUnboundUnbound 

Active-site residues
resource
PDB;1apx & Swiss-prot;P48534 & literature [26], [27], [29], [33]
pdbCatalytic residuesCofactor-binding residuesMain-chain involved in catalysis
           
1apxA01CYS 32;ARG 38;HIS 42
 
TRP 41
1apxB01CYS 32;ARG 38;HIS 42
 
TRP 41
1apxC01CYS 32;ARG 38;HIS 42
 
TRP 41
1apxD01CYS 32;ARG 38;HIS 42
 
TRP 41
1oafA01CYS 32;ARG 38;HIS 42
 
TRP 41
1oagA01CYS 32;ARG 38;HIS 42
 
TRP 41
1v0hX01CYS 32;ARG 38;HIS 42
 
TRP 41
1apxA02ARG 172;ASP 208
HIS 163(Heme binding);THR 164;THR 180;ASN 182;ILE 185;ASP 187(Potassium or Calcium binding)
TRP 179
1apxB02ARG 172;ASP 208
HIS 163(Heme binding);THR 164;THR 180;ASN 182;ILE 185;ASP 187(Potassium or Calcium binding)
TRP 179
1apxC02ARG 172;ASP 208
HIS 163(Heme binding);THR 164;THR 180;ASN 182;ILE 185;ASP 187(Potassium or Calcium binding)
TRP 179
1apxD02ARG 172;ASP 208
HIS 163(Heme binding);THR 164;THR 180;ASN 182;ILE 185;ASP 187(Potassium or Calcium binding)
TRP 179
1oafA02ARG 172;ASP 208
HIS 163(Heme binding);THR 164;THR 180;ASN 182;ILE 185;ASP 187(Potassium or Calcium binding)
TRP 179
1oagA02ARG 172;ASP 208
HIS 163(Heme binding);THR 164;THR 180;ASN 182;ILE 185;ASP 187(Potassium or Calcium binding)
TRP 179
1v0hX02ARG 172;ASP 208
HIS 163(Heme binding);THR 164;THR 180;ASN 182;ILE 185;ASP 187(Potassium or Calcium binding)
TRP 179

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[13]

[19]

[21]p.329-339
[26]

[27]

[29]Scheme 1
[31]Fig.10, p.371-376
[32]p.304-306
[33]Scheme 1, p.32-35
[34]p.8649-8650

references
[1]
PubMed ID1915856
JournalFEBS Lett
Year1991
Volume289
Pages257-9
AuthorsMittler R, Zilinskas BA
TitleMolecular cloning and nucleotide sequence analysis of a cDNA encoding pea cytosolic ascorbate peroxidase.
[2]
PubMed ID1400489
JournalJ Biol Chem
Year1992
Volume267
Pages21802-7
AuthorsMittler R, Zilinskas BA
TitleMolecular cloning and characterization of a gene encoding pea cytosolic ascorbate peroxidase.
[3]
PubMed ID8422923
JournalFEBS Lett
Year1993
Volume315
Pages313-7
AuthorsKubo A, Saji H, Tanaka K, Kondo N
TitleGenomic DNA structure of a gene encoding cytosolic ascorbate peroxidase from Arabidopsis thaliana.
[4]
PubMed ID8006006
JournalJ Biol Chem
Year1994
Volume269
Pages17020-4
AuthorsPatterson WR, Poulos TL
TitleCharacterization and crystallization of recombinant pea cytosolic ascorbate peroxidase.
[5]
PubMed ID7672248
JournalBiochem Soc Trans
Year1995
Volume23
Pages228-32
AuthorsPoulos TL, Patterson WR, Sundaramoorthy M
TitleThe crystal structure of ascorbate and manganese peroxidases: the role of non-haem metal in the catalytic mechanism.
[6]
CommentsX-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS)
Medline ID95217899
PubMed ID7703247
JournalBiochemistry
Year1995
Volume34
Pages4331-41
AuthorsPatterson WR, Poulos TL
TitleCrystal structure of recombinant pea cytosolic ascorbate peroxidase.
Related PDB1apx
Related UniProtKBP48534
[7]
PubMed ID8638916
JournalArch Biochem Biophys
Year1996
Volume328
Pages1-8
AuthorsDalton DA, Diaz del Castillo L, Kahn ML, Joyner SL, Chatfield JM
TitleHeterologous expression and characterization of soybean cytosolic ascorbate peroxidase.
[8]
PubMed ID8766820
JournalFEBS Lett
Year1996
Volume389
Pages153-6
AuthorsMarquez LA, Quitoriano M, Zilinskas BA, Dunford HB
TitleKinetic and spectral properties of pea cytosolic ascorbate peroxidase.
[9]
PubMed ID9291097
JournalBiochem J
Year1997
Volume326
Pages305-10
AuthorsJespersen HM, Kjaersgard IV, Ostergaard L, Welinder KG
TitleFrom sequence analysis of three novel ascorbate peroxidases from Arabidopsis thaliana to structure, function and evolution of seven types of ascorbate peroxidase.
[10]
PubMed ID9346287
JournalEur J Biochem
Year1997
Volume248
Pages347-54
AuthorsHill AP, Modi S, Sutcliffe MJ, Turner DD, Gilfoyle DJ, Smith AT, Tam BM, Lloyd E
TitleChemical, spectroscopic and structural investigation of the substrate-binding site in ascorbate peroxidase.
[11]
PubMed ID9144965
JournalPlant Cell
Year1997
Volume9
Pages627-40
AuthorsKarpinski S, Escobar C, Karpinska B, Creissen G, Mullineaux PM
TitlePhotosynthetic electron transport regulates the expression of cytosolic ascorbate peroxidase genes in Arabidopsis during excess light stress.
[12]
PubMed ID9578600
JournalArch Biochem Biophys
Year1998
Volume353
Pages55-63
AuthorsYoshimura K, Ishikawa T, Nakamura Y, Tamoi M, Takeda T, Tada T, Nishimura K, Shigeoka S
TitleComparative study on recombinant chloroplastic and cytosolic ascorbate peroxidase isozymes of spinach.
[13]
PubMed ID9860877
JournalBiochemistry
Year1998
Volume37
Pages17610-7
AuthorsMandelman D, Jamal J, Poulos TL
TitleIdentification of two electron-transfer sites in ascorbate peroxidase using chemical modification, enzyme kinetics, and crystallography.
[14]
PubMed ID9609702
JournalBiochemistry
Year1998
Volume37
Pages8080-7
AuthorsNissum M, Neri F, Mandelman D, Poulos TL, Smulevich G
TitleSpectroscopic characterization of recombinant pea cytosolic ascorbate peroxidase: similarities and differences with cytochrome c peroxidase.
[15]
PubMed ID9443387
JournalPlanta
Year1998
Volume204
Pages120-6
AuthorsCaldwell CR, Turano FJ, McMahon MB
TitleIdentification of two cytosolic ascorbate peroxidase cDNAs from soybean leaves and characterization of their products by functional expression in E. coli.
[16]
PubMed ID9792095
JournalProtein Sci
Year1998
Volume7
Pages2089-98
AuthorsMandelman D, Schwarz FP, Li H, Poulos TL
TitleThe role of quaternary interactions on the stability and activity of ascorbate peroxidase.
[17]
PubMed ID10217180
JournalElectrophoresis
Year1999
Volume20
Pages630-6
AuthorsKomatsu S, Muhammad A, Rakwal R
TitleSeparation and characterization of proteins from green and etiolated shoots of rice (Oryza sativa L.): towards a rice proteome.
[18]
PubMed ID10729193
JournalArch Biochem Biophys
Year2000
Volume376
Pages82-90
AuthorsTakeda T, Yoshimura K, Yoshii M, Kanahoshi H, Miyasaka H, Shigeoka S
TitleMolecular characterization and physiological role of ascorbate peroxidase from halotolerant Chlamydomonas sp. W80 strain.
[19]
PubMed ID10858284
JournalBiochemistry
Year2000
Volume39
Pages7374-9
AuthorsBursey EH, Poulos TL
TitleTwo substrate binding sites in ascorbate peroxidase: the role of arginine 172.
[20]
PubMed ID11128006
JournalPhilos Trans R Soc Lond B Biol Sci
Year2000
Volume355
Pages1531-40
AuthorsMullineaux P, Ball L, Escobar C, Karpinska B, Creissen G, Karpinski S
TitleAre diverse signalling pathways integrated in the regulation of arabidopsis antioxidant defence gene expression in response to excess excitation energy?
[21]
PubMed ID11192727
JournalSubcell Biochem
Year2000
Volume35
Pages317-49
AuthorsRaven EL
TitlePeroxidase-catalyzed oxidation of ascorbate. Structural, spectroscopic and mechanistic correlations in ascorbate peroxidase.
[22]
PubMed ID11361125
JournalArch Biochem Biophys
Year2001
Volume388
Pages100-12
AuthorsBattistuzzi G, D'Onofrio M, Loschi L, Sola M
TitleIsolation and characterization of two peroxidases from Cucumis sativus.
[23]
PubMed ID11356136
JournalBiochem Soc Trans
Year2001
Volume29
Pages105-11
AuthorsLloyd Raven E, Celik A, Cullis PM, Sangar R, Sutcliffe MJ
TitleEngineering the active site of ascorbate peroxidase.
[24]
PubMed ID11696974
JournalDNA Seq
Year2001
Volume11
Pages475-84
AuthorsKim IJ, Lee BH, Jo J, Chung WI
TitleSequence variability of nine cytosolic ascorbate peroxidases in polyploid strawberry.
[25]
PubMed ID11121105
JournalEur J Biochem
Year2001
Volume268
Pages78-85
AuthorsCelik A, Cullis PM, Sutcliffe MJ, Sangar R, Raven EL
TitleEngineering the active site of ascorbate peroxidase.
[26]
PubMed ID11358529
JournalEur J Biochem
Year2001
Volume268
Pages3091-8
AuthorsHiner AN, Martinez JI, Arnao MB, Acosta M, Turner DD, Lloyd Raven E, Rodriguez-Lopez JN
TitleDetection of a tryptophan radical in the reaction of ascorbate peroxidase with hydrogen peroxide.
[27]
PubMed ID12427040
JournalBiochemistry
Year2002
Volume41
Pages13774-81
AuthorsLad L, Mewies M, Raven EL
TitleSubstrate binding and catalytic mechanism in ascorbate peroxidase: evidence for two ascorbate binding sites.
[28]
PubMed ID12506974
JournalBiosci Biotechnol Biochem
Year2002
Volume66
Pages2367-75
AuthorsKitajima S, Ueda M, Sano S, Miyake C, Kohchi T, Tomizawa K, Shigeoka S, Yokota A
TitleStable form of ascorbate peroxidase from the red alga Galdieria partita similar to both chloroplastic and cytosolic isoforms of higher plants.
[29]
PubMed ID12084058
JournalEur J Biochem
Year2002
Volume269
Pages3182-92
AuthorsLad L, Mewies M, Basran J, Scrutton NS, Raven EL
TitleRole of histidine 42 in ascorbate peroxidase. Kinetic analysis of the H42A and H42E variants.
[30]
PubMed ID12966073
JournalJ Biochem (Tokyo)
Year2003
Volume134
Pages239-44
AuthorsWada K, Tada T, Nakamura Y, Ishikawa T, Yabuta Y, Yoshimura K, Shigeoka S, Nishimura K
TitleCrystal structure of chloroplastic ascorbate peroxidase from tobacco plants and structural insights into its instability.
[31]
PubMed ID12964833
JournalNat Prod Rep
Year2003
Volume20
Pages367-81
AuthorsRaven EL
TitleUnderstanding functional diversity and substrate specificity in haem peroxidases: what can we learn from ascorbate peroxidase?
[32]
PubMed ID12640445
JournalNat Struct Biol
Year2003
Volume10
Pages303-7
AuthorsSharp KH, Mewies M, Moody PC, Raven EL
TitleCrystal structure of the ascorbate peroxidase-ascorbate complex.
Related PDB1oaf,1oag
[33]
PubMed ID15777010
JournalBiochem Soc Symp
Year2004
Volume(71)
Pages27-38
AuthorsRaven EL, Lad L, Sharp KH, Mewies M, Moody PC
TitleDefining substrate specificity and catalytic mechanism in ascorbate peroxidase.
[34]
PubMed ID15236572
JournalBiochemistry
Year2004
Volume43
Pages8644-51
AuthorsSharp KH, Moody PC, Brown KA, Raven EL
TitleCrystal structure of the ascorbate peroxidase-salicylhydroxamic acid complex.
Related PDB1v0h

comments
According to the literature, this enzyme catalyzes three successive reactions:
(A) Enzyme + H2O2 => Enzyme Intermediate I + H2O
(B) Enzyme Intermediate I + AH => Enzyme Intermediate II + A*
(C) Enzyme Intermediate II + AH => Enzyme + A* + H2O
Here, AH is substrate, ascorbate, whereas A* is a radical form of ascorbate. Enzyme Intermediate I contains oxyferryl [Fe(IV)=O].

createdupdated
2004-05-112009-02-26


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 2008)
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)
Funded by Grant-in-Aid for Publication of Scientific Research Results/Japan Society for the Promotion of Science (JSPS) (April 2012 - March 2013)
Supported by the commission for the Development of Artificial Gene Synthesis Technology for Creating Innovative Biomaterial from the Ministry of Economy, Trade and Industry (METI) (October 2012 - )
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