Crystallization (Comment) | Organism |
---|---|
wild type holo and apoenzymes and mutant enzymes C151S, C151G and C151S/H178N in complex with NAD+ and D-glyceraldehyde 3-phosphate, hanging drop vapor diffusion method, the wild-type holoenzyme and apoenzyme are crystallized from 0.1 M Tris-HCl, pH 8.5, and 20% (w/v) PEG 4000 at 4°C and from 0.1 M Tris-HCl, pH 8.2, and 32% (w/v) PEG 3350 at 25°C, respectively. Crystals of active-site mutants in complex with NAD+ (C151S, C151G, and C151S/H178N) grow at 25°C from 0.1 M Tris-HCl, pH 8.5, and 28% (w/v) PEG 4000, from 0.1 M Tris-HCl, pH 8.2, and 30% (w/v) PEG 4000, and from 0.1 M Tris-HCl, pH 8.5, and 30% (w/v) PEG 4000, respectively, while mutant H178N in complex with NAD+ crystallizes from 0.1 M Tris-HCl, pH 8.2, and 30% (w/v) PEG 4000 at 4°C | Staphylococcus aureus |
Protein Variants | Comment | Organism |
---|---|---|
C151/H178N | the rate of the forward reaction is decreased by 47000 times compared to the wild type enzyme with similar affinity for the substrate and the coenzyme | Staphylococcus aureus |
C151G | the mutant is completely inactive | Staphylococcus aureus |
C151S | the mutant shows drastically reduced kcat values compared to the wild type enzyme | Staphylococcus aureus |
H178N | the mutant shows no significant difference in the Km values of D-glyceraldehyde 3-phosphate, phosphate, and NAD+ compared to the wild type enzyme, however, the mutation results in the reduction of kcat of oxidative phosphorylation by 1400times | Staphylococcus aureus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.31 | - |
D-glyceraldehyde 3-phosphate | wild type enzyme, pH 8.7, 25°C | Staphylococcus aureus | |
0.32 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme H178N, pH 8.7, 25°C | Staphylococcus aureus | |
0.34 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme C151S, pH 8.7, 25°C | Staphylococcus aureus | |
0.34 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme C151S/H178N, pH 8.7, 25°C | Staphylococcus aureus | |
2.54 | - |
phosphate | wild type enzyme, pH 8.7, 25°C | Staphylococcus aureus | |
3.03 | - |
phosphate | mutant enzyme H178N, pH 8.7, 25°C | Staphylococcus aureus | |
3.09 | - |
phosphate | mutant enzyme C151S, pH 8.7, 25°C | Staphylococcus aureus | |
3.68 | - |
phosphate | mutant enzyme C151S/H178N, pH 8.7, 25°C | Staphylococcus aureus | |
312 | - |
NAD+ | mutant enzyme C151S/H178N, pH 8.7, 25°C | Staphylococcus aureus | |
316 | - |
NAD+ | wild type enzyme, pH 8.7, 25°C | Staphylococcus aureus | |
320 | - |
NAD+ | mutant enzyme H178N, pH 8.7, 25°C | Staphylococcus aureus | |
322 | - |
NAD+ | mutant enzyme C151S, pH 8.7, 25°C | Staphylococcus aureus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | Q6GIL8 | - |
- |
Staphylococcus aureus MRSA252 | Q6GIL8 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | - |
Staphylococcus aureus | 3-phospho-D-glyceroyl phosphate + NADH | - |
? | |
D-glyceraldehyde 3-phosphate + phosphate + NAD+ | - |
Staphylococcus aureus MRSA252 | 3-phospho-D-glyceroyl phosphate + NADH | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | x-ray crystallography | Staphylococcus aureus |
Synonyms | Comment | Organism |
---|---|---|
GAPDH1 | - |
Staphylococcus aureus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
- |
Staphylococcus aureus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.002 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme C151S/H178N, pH 8.7, 25°C | Staphylococcus aureus | |
0.05 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme H178N, pH 8.7, 25°C | Staphylococcus aureus | |
0.09 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme C151S, pH 8.7, 25°C | Staphylococcus aureus | |
70 | - |
D-glyceraldehyde 3-phosphate | wild type enzyme, pH 8.7, 25°C | Staphylococcus aureus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.7 | - |
- |
Staphylococcus aureus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Staphylococcus aureus |