Cloned (Comment) | Organism |
---|---|
subcloning in Escherichia coli strain XL1-blue, expression of wild-type and mutant enzymes in enzyme-deficient Saccharomyces cerevisiae strain BJ268 | Vigna radiata |
Protein Variants | Comment | Organism |
---|---|---|
A226S | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
A238S | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Vigna radiata |
E225A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Vigna radiata |
F224A | site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme | Vigna radiata |
F240A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Vigna radiata |
G221A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
G222A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
G229A | site-directed mutagenesis, the mutant shows 50% reduced activity compared to the wild-type enzyme | Vigna radiata |
G231A | site-directed mutagenesis, the mutant shows 50% reduced activity compared to the wild-type enzyme | Vigna radiata |
G233A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
G234A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
G241A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
I227A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
L223A | site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme | Vigna radiata |
L232A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Vigna radiata |
L239A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
M237A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Vigna radiata |
R242A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
S235A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
S236A | site-directed mutagenesis, the mutant shows slightly reduced activity compared to the wild-type enzyme | Vigna radiata |
T228A | site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme | Vigna radiata |
Y230A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Vigna radiata |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | - |
Vigna radiata | |
F- | - |
Vigna radiata | |
Na+ | - |
Vigna radiata |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes | Vigna radiata | |
0.008 | - |
diphosphate | pH 8.0, 37°C, mutant Y230A | Vigna radiata | |
0.016 | - |
diphosphate | pH 8.0, 37°C, mutant G231A | Vigna radiata | |
0.022 | - |
diphosphate | pH 8.0, 37°C, mutant R242A | Vigna radiata | |
0.035 | - |
diphosphate | pH 8.0, 37°C, mutant L232A | Vigna radiata | |
0.054 | - |
diphosphate | pH 8.0, 37°C, mutant E225A | Vigna radiata | |
0.068 | - |
diphosphate | pH 8.0, 37°C, mutant G229A | Vigna radiata | |
0.07 | - |
diphosphate | pH 8.0, 37°C, mutant G234A | Vigna radiata | |
0.074 | - |
diphosphate | pH 8.0, 37°C, mutant G233A | Vigna radiata | |
0.099 | - |
diphosphate | pH 8.0, 37°C, mutant F240A | Vigna radiata | |
0.105 | - |
diphosphate | pH 8.0, 37°C, mutant S236A | Vigna radiata | |
0.112 | - |
diphosphate | pH 8.0, 37°C, mutant M237A | Vigna radiata | |
0.14 | - |
diphosphate | pH 8.0, 37°C, mutant I227A | Vigna radiata | |
0.146 | - |
diphosphate | pH 8.0, 37°C, mutant T228A | Vigna radiata | |
0.148 | - |
diphosphate | pH 8.0, 37°C, mutant G221A | Vigna radiata | |
0.154 | - |
diphosphate | pH 8.0, 37°C, wild-type enzyme | Vigna radiata | |
0.185 | - |
diphosphate | pH 8.0, 37°C, mutant A226S | Vigna radiata | |
0.196 | - |
diphosphate | pH 8.0, 37°C, mutant A238S | Vigna radiata | |
0.199 | - |
diphosphate | pH 8.0, 37°C, mutant S235A | Vigna radiata | |
0.221 | - |
diphosphate | pH 8.0, 37°C, mutant L239A | Vigna radiata | |
0.305 | - |
diphosphate | pH 8.0, 37°C, mutant F224A | Vigna radiata | |
0.318 | - |
diphosphate | pH 8.0, 37°C, mutant G222A | Vigna radiata | |
0.332 | - |
diphosphate | pH 8.0, 37°C, mutant G241A | Vigna radiata | |
0.438 | - |
diphosphate | pH 8.0, 37°C, mutant L223A | Vigna radiata |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
vacuolar membrane | the enzyme contains several transmembrane segments | Vigna radiata | 5774 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | required, highly activating | Vigna radiata | |
Mg2+ | required | Vigna radiata |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
81000 | - |
- |
Vigna radiata |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
diphosphate + H2O | Vigna radiata | - |
2 phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vigna radiata | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
diphosphate + H2O | - |
Vigna radiata | 2 phosphate | - |
? | |
additional information | the enzyme acts as diphosphate-dependent H+-translocation pump | Vigna radiata | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | the homodimer contains several transmembrane segments, the conserved transmembrane segment 5 at the N-terminal side near the putative substrate binding loop contains the GYG motif which is important in maintaining structure and function of the enzyme, overview | Vigna radiata |
Synonyms | Comment | Organism |
---|---|---|
V-PPase | - |
Vigna radiata |
vacuolar H+-pyrophosphatase | - |
Vigna radiata |
vacuolar H+-translocating inorganic pyrophosphatase | - |
Vigna radiata |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Vigna radiata |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | 8.1 | pH optima of mutant enzymes, overview | Vigna radiata |
8 | - |
pH optimum of the wild-type enzyme | Vigna radiata |