EzCatDB: T00237
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DB codeT00237
CATH domainDomain 13.40.50.970 : Rossmann foldCatalytic domain
Domain 23.40.50.1220 : Rossmann foldCatalytic domain
Domain 33.40.50.970 : Rossmann foldCatalytic domain
E.C.1.2.3.3
CSA1pow
MACiEM0274

CATH domainRelated DB codes (homologues)
3.40.50.1220 : Rossmann foldM00162,M00161,T00230
3.40.50.970 : Rossmann foldT00210,T00230

Enzyme Name
UniProtKBKEGG

P37063
Protein namePyruvate oxidasepyruvate oxidase
pyruvic oxidase
phosphate-dependent pyruvate oxidase
SynonymsEC 1.2.3.3
Pyruvic oxidase
POX
RefSeqYP_004891064.1 (Protein)
NC_004567.2 (DNA/RNA sequence)
PfamPF02775 (TPP_enzyme_C)
PF00205 (TPP_enzyme_M)
PF02776 (TPP_enzyme_N)
[Graphical view]

KEGG pathways
MAP codePathways
MAP00620Pyruvate metabolism

UniProtKB:Accession NumberP37063
Entry namePOXB_LACPL
ActivityPyruvate + phosphate + O(2) = acetyl phosphate + CO(2) + H(2)O(2).
SubunitHomotetramer.
Subcellular location
CofactorBinds 1 FAD per subunit.,Binds 1 magnesium ion per subunit.,Binds 1 thiamine pyrophosphate per subunit.

Compound table: links to PDB-related databases & PoSSuM

CofactorsSubstratesProducts
KEGG-idC00016C00068C00305C00022C00009C00007C00227C00011C00027
CompoundFADThiamine diphosphateMagnesiumPyruvateOrthophosphateO2Acetyl phosphateCO2H2O2
Typeamide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carbohydrate,nucleotideamine group,aromatic ring (with nitrogen atoms),phosphate group/phosphate iondivalent metal (Ca2+, Mg2+)carbohydrate,carboxyl groupphosphate group/phosphate ionotherscarbohydrate,phosphate group/phosphate ionothersothers
ChEBI16238
9532
18420
32816
26078
27140
26689
15379
15350
16526
16240
PubChem643975
1132
888
1060
22486802
1004
977
186
280
784
22326046
                 
1powA01UnboundUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1powB01UnboundUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1poxA01UnboundUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1poxB01UnboundUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1y9dA01UnboundUnboundUnboundUnboundAnalogue:SO4UnboundUnboundUnboundUnbound
1y9dB01UnboundUnboundUnboundUnboundAnalogue:SO4UnboundUnboundUnboundUnbound
1y9dC01UnboundUnboundUnboundUnboundAnalogue:SO4UnboundUnboundUnboundUnbound
1y9dD01UnboundUnboundUnboundUnboundAnalogue:SO4UnboundUnboundUnboundUnbound
1powA02Bound:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1powB02Bound:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1poxA02Bound:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1poxB02Bound:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1y9dA02Analogue:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1y9dB02Analogue:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1y9dC02Analogue:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1y9dD02Analogue:FADUnboundUnboundUnboundUnboundUnboundUnboundUnboundUnbound
1powA03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound
1powB03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound
1poxA03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound
1poxB03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound
1y9dA03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound
1y9dB03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound
1y9dC03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound
1y9dD03UnboundBound:TPPBound:_MGUnboundUnboundUnboundUnboundUnboundUnbound

Active-site residues
resource
Swiss-prot;P37063 & literature [5], [8], [10]
pdbCatalytic residuesCofactor-binding residuescomment
           
1powA01GLU 59
 
 
1powB01GLU 59
 
 
1poxA01GLU 59
 
mutant P178S, S188N
1poxB01GLU 59
 
mutant P178S, S188N
1y9dA01GLU 59
 
 
1y9dB01GLU 59
 
 
1y9dC01GLU 59
 
 
1y9dD01GLU 59
 
 
1powA02ARG 264
 
 
1powB02ARG 264
 
 
1poxA02ARG 264
 
 
1poxB02ARG 264
 
 
1y9dA02ARG 264
 
mutant V265A
1y9dB02ARG 264
 
mutant V265A
1y9dC02ARG 264
 
mutant V265A
1y9dD02ARG 264
 
mutant V265A
1powA03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
 
1powB03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
 
1poxA03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
mutant A458V
1poxB03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
mutant A458V
1y9dA03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
 
1y9dB03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
 
1y9dC03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
 
1y9dD03PHE 479;GLU 483
ASP 447;ASN 474;GLN 476(Magnesium binding)
 

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[3]p.967
[4]

[5]p.331-333
[8]SCHEME 1, p.12933-12934
[10]Scheme 1, Scheme 3, p.10754

references
[1]
PubMed ID4896238
JournalAnnu Rev Biochem
Year1969
Volume38
Pages213-40
AuthorsKrampitz LO
TitleCatalytic functions of thiamin diphosphate.
[2]
PubMed ID2037573
JournalJ Biol Chem
Year1991
Volume266
Pages10168-73
AuthorsBertagnolli BL, Hager LP
TitleActivation of Escherichia coli pyruvate oxidase enhances the oxidation of hydroxyethylthiamin pyrophosphate.
[3]
CommentsX-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS)
Medline ID93174262
PubMed ID8438155
JournalScience
Year1993
Volume259
Pages965-7
AuthorsMuller YA, Schulz GE
TitleStructure of the thiamine- and flavin-dependent enzyme pyruvate oxidase.
Related UniProtKBP37063
[4]
PubMed ID8069629
JournalStructure
Year1993
Volume1
Pages95-103
AuthorsMuller YA, Lindqvist Y, Furey W, Schulz GE, Jordan F, Schneider G
TitleA thiamin diphosphate binding fold revealed by comparison of the crystal structures of transketolase, pyruvate oxidase and pyruvate decarboxylase.
[5]
CommentsX-RAY CRYSTALLOGRAPHY (2.5 AND 2.1 ANGSTROMS) OF WILD-TYPE AND MUTANT
Medline ID94194507
PubMed ID8145244
JournalJ Mol Biol
Year1994
Volume237
Pages315-35
AuthorsMuller YA, Schumacher G, Rudolph R, Schulz GE
TitleThe refined structures of a stabilized mutant and of wild-type pyruvate oxidase from Lactobacillus plantarum.
Related PDB1pow,1pox
Related UniProtKBP37063
[6]
PubMed ID9305946
JournalBiochemistry
Year1997
Volume36
Pages11564-73
AuthorsChang YY, Cronan JE Jr
TitleSulfhydryl chemistry detects three conformations of the lipid binding region of Escherichia coli pyruvate oxidase.
[7]
PubMed ID9655909
JournalBiochim Biophys Acta
Year1998
Volume1385
Pages221-8
AuthorsHubner G, Tittmann K, Killenberg-Jabs M, Schaffner J, Spinka M, Neef H, Kern D, Kern G, Schneider G, Wikner C, Ghisla S
TitleActivation of thiamin diphosphate in enzymes.
[8]
PubMed ID9582325
JournalJ Biol Chem
Year1998
Volume273
Pages12929-34
AuthorsTittmann K, Proske D, Spinka M, Ghisla S, Rudolph R, Hubner G, Kern G
TitleActivation of thiamin diphosphate and FAD in the phosphatedependent pyruvate oxidase from Lactobacillus plantarum.
[9]
CommentsReview in a book; Academic Press Ltd., San Diego
JournalComprehensive Biological Catalysis (Editor: Sinnott M)
Year1998
Volume
Pages217-66
AuthorsSchowen RL
TitleThiamin-dependent Enzymes
[10]
PubMed ID10978159
JournalBiochemistry
Year2000
Volume39
Pages10747-54
AuthorsTittmann K, Golbik R, Ghisla S, Hubner G
TitleMechanism of elementary catalytic steps of pyruvate oxidase from Lactobacillus plantarum.
[11]
PubMed ID10617618
JournalJ Biol Chem
Year2000
Volume275
Pages297-302
AuthorsSvergun DI, Petoukhov MV, Koch MH, Konig S
TitleCrystal versus solution structures of thiamine diphosphate-dependent enzymes.
[12]
PubMed ID11170450
JournalBiochemistry
Year2001
Volume40
Pages1248-56
AuthorsMarchal D, Pantigny J, Laval JM, Moiroux J, Bourdillon C
TitleRate constants in two dimensions of electron transfer between pyruvate oxidase, a membrane enzyme, and ubiquinone (coenzyme Q8), its water-insoluble electron carrier.
[13]
PubMed ID11514662
JournalProtein Sci
Year2001
Volume10
Pages1712-28
AuthorsDym O, Eisenberg D
TitleSequence-structure analysis of FAD-containing proteins.
[14]
PubMed ID15794646
JournalBiochemistry
Year2005
Volume44
Pages5086-94
AuthorsWille G, Ritter M, Weiss MS, Konig S, Mantele W, Hubner G
TitleThe role of Val-265 for flavin adenine dinucleotide (FAD) binding in pyruvate oxidase: FTIR, kinetic, and crystallographic studies on the enzyme variant V265A.
Related PDB1y9d

comments
Although this enzyme binds magnesium ion, it is not involved in catalysis.
According to the literature [5], this enzyme catalyzes several reactions (Eq.1-Eq.4) as follwos:
(Eq.1) Pyruvate + ThDP-E-FAD(ox) = Oxyethyl-ThDP-E-FAD(ox) + CO2
(A) Addition of ThDP to Pyruvate
(B) Elimination of carboxylate (CO2), leading to Oxyethyl-ThDP Intermediate
(Eq.2) Oxyethyl-ThDP-E-FAD(ox) = Acetyl-ThDP-E-FAD(red)
(C) Hydride transfer from Oxyethyl-ThDP intermediate to FAD(ox)
(Eq.3) Acetyl-ThDP-E-FAD(red) + O2 = Acetyl-ThDP-E-FAD(ox) + H2O2
(D) Hydride transfer from FAD(red) to O2 (probably) Or oxygenation of FAD
(Eq.4) Acetyl-ThDP-E-FAD(ox) + Phosphate = Acetylphosphate + ThDP-E-FAD(ox)
(E) Phosphorolysis of Acetyl-ThDP intermediate

createdupdated
2004-03-242009-02-26


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