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Literature summary for 1.8.3.2 extracted from

  • Wang, W.; Winther, J.R.; Thorpe, C.
    Erv2p: characterization of the redox behavior of a yeast sulfhydryl oxidase (2007), Biochemistry, 46, 3246-3254.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
truncated version of Erv2p, without the N-terminal 34 residues, cloned into a pET24(a+) plasmid carrying kanamycin resistance. Amplified in DH5alpha Escherichia coli strain. Expressed from the Escherichia coli strain BL21(DE3) or BL21(DE3 star) Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
Zn2+ upon coordination with Zn2+, full reduction of Erv2p requires 6 electrons. Strongly inhibits Erv2p when assayed using tris(2-carboxyethyl)phosphine as the reducing substrate of the oxidase. Restoration of 1 mM EDTA effects rapid recovery of 80% of the original activity Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
11.3
-
dithiothreitol
-
Saccharomyces cerevisiae

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
Saccharomyces cerevisiae 16020
-

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
gel filtration Saccharomyces cerevisiae

Storage Stability

Storage Stability Organism
-20°C, Tris buffer, pH 7.5, 1 mM EDTA, more than 1 year Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
bis-(2-mercaptoethyl)sulfone + O2
-
Saccharomyces cerevisiae ? + H2O
-
?
dithiothreitol + O2
-
Saccharomyces cerevisiae ? + H2O
-
?
additional information Erv2p is a modest catalyst of disulfide bond formation. None of the monothiols (at 10 mM), including beta-mercaptoethanol, N-acetylcysteamine, reduced glutathione and CoASH, prove detectable substrates of the yeast oxidase at pH 7.5. In contrast, dithiols are significant substrates Saccharomyces cerevisiae ?
-
?
reduced lysozyme + O2
-
Saccharomyces cerevisiae ? + H2O
-
?
RNasered + O2
-
Saccharomyces cerevisiae ? + H2O
-
?
thioredoxin + O2
-
Saccharomyces cerevisiae thioredoxin disulfide + H2O2
-
?
tris(2-carboxyethyl)-phosphine + O2
-
Saccharomyces cerevisiae ? + H2O
-
?
Trx Escherichia coli + O2
-
Saccharomyces cerevisiae ? + H2O
-
?

Synonyms

Synonyms Comment Organism
ERv2p
-
Saccharomyces cerevisiae
sulfhydryl oxidase
-
Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.43
-
dithiothreitol
-
Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
FAD dithionite and photochemical reductions of Erv2p show full reduction of the flavin cofactor after the addition of 4 electrons with a midpoint potential of -200 mV at pH 7.5. No charge-transfer complex between a proximal thiolate and the oxidized flavin Saccharomyces cerevisiae