Cloned (Comment) | Organism |
---|---|
expression of His6-tagged enzyme in Escherichia coli | Saccharomyces cerevisiae |
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 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 | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrial intermembrane space | - |
Saccharomyces cerevisiae | 5758 | - |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
flavoprotein | - |
Saccharomyces cerevisiae |
Purification (Comment) | Organism |
---|---|
recobinant His6-tagged enzyme from Escherichia coli by nickel affinity chromatography and gel filtration | Saccharomyces cerevisiae |
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 | 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 | Comment | Organism |
---|---|---|
Erv1p | - |
Saccharomyces cerevisiae |
sulfhydryl oxidase | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Saccharomyces cerevisiae |
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 | Comment | Organism | Structure |
---|---|---|---|
FAD | dependent on. The three disulfides of the enzyme are not essential for FAD binding | Saccharomyces cerevisiae |
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 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 |