Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Arabidopsis thaliana |
Protein Variants | Comment | Organism |
---|---|---|
C149S | the mutant shows 92% activity compared to the wild type enzyme | Arabidopsis thaliana |
C149S | about 1% of wild-type activity, residue Cys149 is both essential for catalysis and the only accessible cysteine | Arabidopsis thaliana |
C153S | the mutant shows 0.7% activity compared to the wild type enzyme | Arabidopsis thaliana |
C153S | residue Cys153 has apparently no role in catalysis, in spite of the proximity in space with catalytic Cys149 | Arabidopsis thaliana |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
H2O2 | inhibits enzyme activity by converting the thiolate of Cys149 into irreversibly oxidized forms, -SO2- and SO3- via a labile sulfenate intermediate SO?. Reduced glutathione prevents this irreversible process by reacting with Cys149 sulfenates to give rise to a mixed disulfide. Glutathionylated enzyme can be fully reactivated either by cytosolic glutaredoxin, via a glutathione-dependent monothiol mechanism, or, less efficiently, by cytosolic thioredoxins physiologically reduced by NADPH:thioredoxin reductase | Arabidopsis thaliana |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytosol | - |
Arabidopsis thaliana | 5829 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
x * 40000, SDS-PAGE | Arabidopsis thaliana |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | Arabidopsis thaliana | - |
3-phospho-D-glyceroyl phosphate + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | - |
- |
- |
Arabidopsis thaliana | P25858 | - |
- |
Oxidation Stability | Organism |
---|---|
isoform GapC1 is sensitive to oxidation by 0.05 mM H2O2, which appears to inhibit enzyme activity by converting the thiolate of Cys149 (S) into irreversible oxidized forms (SO2 and SO3) via a labile sulfenate intermediate (SO) | Arabidopsis thaliana |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.4 | - |
mutant enzyme C149S, 50 mM Tris/HCl (pH 7.5), at 25°C | Arabidopsis thaliana |
0.4 | - |
mutant C149S, pH 7.5, 25°C | Arabidopsis thaliana |
51.1 | - |
mutant enzyme C153S, 50 mM Tris/HCl (pH 7.5), at 25°C | Arabidopsis thaliana |
51.1 | - |
mutant C153S, pH 7.5, 25°C | Arabidopsis thaliana |
55.5 | - |
wild-type, pH 7.5, 25°C | Arabidopsis thaliana |
55.5 | - |
His-tagged wild type enzyme, 50 mM Tris/HCl (pH 7.5), at 25°C | Arabidopsis thaliana |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | - |
Arabidopsis thaliana | 3-phospho-D-glyceroyl phosphate + NADH | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | - |
Arabidopsis thaliana | 3-phospho-D-glyceroyl phosphate + NADH + H+ | - |
? | |
additional information | enzymatic activity strictly depends on residue Cys149. Catalytic Cys149 is the only solvent-exposed cysteine of the protein and its thiol is relatively acidic, pKa =5.7 | Arabidopsis thaliana | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 40000, SDS-PAGE | Arabidopsis thaliana |
Synonyms | Comment | Organism |
---|---|---|
At3g04120 | - |
Arabidopsis thaliana |
GapC1 | - |
Arabidopsis thaliana |
GapC1 | isoform | Arabidopsis thaliana |
GAPDH | - |
Arabidopsis thaliana |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | dependent on, the activity with NADPH is negligible (0.15% of the NADH-dependent specific activity) | Arabidopsis thaliana |