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Literature summary for 1.8.3.2 extracted from

  • Ang, S.K.; Lu, H.
    Deciphering structural and functional roles of individual disulfide bonds of the mitochondrial sulfhydryl oxidase Erv1p (2009), J. Biol. Chem., 284, 28754-28761.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of His6-tagged enzyme in Escherichia coli Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
C130S/C133S site-directed mutagenesis, the active site mutant shows no or very little activity, and the mutant shows a shifted protein-bound FAD spectrum compared to the wild-type enzyme Erv1p, the active site disulfide is located proximal to the isoalloxazine ring of FADa nd the mutation changes bound-FAD absorption slightly, the mutant is active in presence of DTT, but not with tris(2-carboxyethyl)phosphine Saccharomyces cerevisiae
C159S/C176S site-directed mutagenesis, the mutant shows the same protein-bound FAD spectrum as the wild-type enzyme Erv1p Saccharomyces cerevisiae
C30S/C33S site-directed mutagenesis, the mutant shows the same protein-bound FAD spectrum as the wild-type enzyme Erv1p Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information oxygen consumption kinetic parameters for the WT and Erv1p mutants, overview Saccharomyces cerevisiae
0.018
-
O2 25°C, recombinant mutant C159S/C176S, in presence of 10 3.5 mM tris(2-carboxyethyl)phosphine Saccharomyces cerevisiae
0.027
-
O2 25°C, recombinant wild-type enzyme, in presence of 3.5 mM tris(2-carboxyethyl)phosphine Saccharomyces cerevisiae
0.057
-
O2 25°C, recombinant wild-type enzyme, in presence of 10 mM DTT Saccharomyces cerevisiae
0.062
-
O2 25°C, recombinant mutant C30S/C33S, in presence of 10 mM DTT Saccharomyces cerevisiae
0.087
-
O2 25°C, recombinant mutant C159S/C176S, in presence of 10 mM DTT Saccharomyces cerevisiae

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrial intermembrane space
-
Saccharomyces cerevisiae 5758
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Saccharomyces cerevisiae enzyme regulation, overview ?
-
?
protein Mia40 + O2 Saccharomyces cerevisiae
-
protein Mia40 disulfide + H2O
-
?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
flavoprotein
-
Saccharomyces cerevisiae

Purification (Commentary)

Purification (Comment) Organism
recobinant His6-tagged enzyme from Escherichia coli by nickel affinity chromatography and gel filtration Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information enzyme regulation, overview Saccharomyces cerevisiae ?
-
?
additional information Erv1p contains three conserved disulfide bonds arranged in two CXXC motifs and one CX16C motif, the CX16C disulfide plays an important role in stabilizing the folding of Erv1p, both CXXC disulfides are required for Erv1 oxidase activity, but none of the disulfide is essential for FAD binding, overview Saccharomyces cerevisiae ?
-
?
protein Mia40 + O2
-
Saccharomyces cerevisiae protein Mia40 disulfide + H2O
-
?
protein Mia40 + O2 recombinantly expressed substrate amino acids 284-403, which is the C-terminal domain of Mia40, electron transfer between the shuttle and active site disulfides of Erv1p. Both intersubunit and intermolecular electron transfer can occur, overview Saccharomyces cerevisiae protein Mia40 disulfide + H2O
-
?

Subunits

Subunits Comment Organism
More Erv1p contains three conserved disulfide bonds arranged in two CXXC motifs and one CX16C motif in the highly conserved central catalytic core. The CX16C disulfide plays an important role in stabilizing the folding of Erv1p, both CXXC disulfides are required for Erv1 oxidase activity, but none of the disulfide is essential for FAD binding, overview Saccharomyces cerevisiae

Synonyms

Synonyms Comment Organism
Erv1p
-
Saccharomyces cerevisiae
sulfhydryl oxidase
-
Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.7
-
O2 25°C, recombinant mutant C159S/C176S, in presence of 10 3.5 mM tris(2-carboxyethyl)phosphine Saccharomyces cerevisiae
0.8
-
O2 25°C, recombinant mutant C159S/C176S, in presence of 10 mM DTT Saccharomyces cerevisiae
1.1
-
O2 25°C, recombinant wild-type enzyme, in presence of 3.5 mM tris(2-carboxyethyl)phosphine Saccharomyces cerevisiae
1.3
-
O2 25°C, recombinant wild-type enzyme, in presence of 10 mM DTT Saccharomyces cerevisiae
1.5
-
O2 25°C, recombinant mutant C30S/C33S, in presence of 10 mM DTT Saccharomyces cerevisiae

Cofactor

Cofactor Comment Organism Structure
FAD dependent on. The three disulfides of the enzyme are not essential for FAD binding Saccharomyces cerevisiae

General Information

General Information Comment Organism
physiological function Erv1p is a FAD-dependent sulfhydryl oxidase and is an essential component of the redox regulated Mia40/Erv1 import and assembly pathway used by many of the cysteine-containing intermembrane space proteins Saccharomyces cerevisiae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0023
-
O2 25°C, recombinant wild-type enzyme, in presence of 10 mM DTT Saccharomyces cerevisiae
0.0024
-
O2 25°C, recombinant mutant C30S/C33S, in presence of 10 mM DTT Saccharomyces cerevisiae
0.0039
-
O2 25°C, recombinant mutant C159S/C176S, in presence of 10 3.5 mM tris(2-carboxyethyl)phosphine Saccharomyces cerevisiae
0.0041
-
O2 25°C, recombinant wild-type enzyme, in presence of 3.5 mM tris(2-carboxyethyl)phosphine Saccharomyces cerevisiae
0.0093
-
O2 25°C, recombinant mutant C159S/C176S, in presence of 10 mM DTT Saccharomyces cerevisiae