Activating Compound | Comment | Organism | Structure |
---|---|---|---|
D-fructose 6-phosphate | a slight increase in enzyme activity is observed in the presence of D-fructose 6-phosphate and coenzyme A (1.1fold) | Sulfurisphaera tokodaii | |
D-glucose 1-phosphate | activity increases by 1.4fold with increasing D-glucose 1-phosphate concentration from 0 to 0.1 mM. The enzyme shows K-type allosterism in which the positive cooperativity is abolished with concomitant activation by glucose 1-phosphate | Sulfurisphaera tokodaii | |
D-glucose 1-phosphate | D-glucose 1-phosphate does not influence the affinity for DL-glyceraldehyde-3-phosphate; but increases the turnover severalfold | Sulfurisphaera tokodaii | |
additional information | ATP, ADP, AMP, D-glucose 6-phosphate and galactose 1-phosphate exhibit no effect on enzyme activity | Sulfurisphaera tokodaii |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli C43 (DE3) cells | Sulfurisphaera tokodaii |
expression in Escherichia coli | Sulfurisphaera tokodaii |
Protein Variants | Comment | Organism |
---|---|---|
A198S/S199I | the catalytic efficiency with NADP+ decreases while that with NAD+ increases by 2.5fold. Substitutions reduces the NADP/NAD preference ratio by more than 50% | Sulfurisphaera tokodaii |
A198S/S199I | the mutant shows reduced catalytic efficiency with NADP+ and increased catalytic efficiency with NAD+ as compared to the wild type enzyme | Sulfurisphaera tokodaii |
S199I | the catalytic efficiency (kcat/Km) with NADP+ decreases by 0.5fold while that with NAD+ remains unchanged. Substitution reduces the NADP/NAD preference ratio by more than 50% | Sulfurisphaera tokodaii |
S199I | the mutant shows reduced catalytic efficiency with NADP+ and increased catalytic efficiency with NAD+ as compared to the wild type enzyme | Sulfurisphaera tokodaii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no effect: ADP, AMP, D-glucose 6-phosphate and D-galactose 1-phosphate | Sulfurisphaera tokodaii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.022 | - |
NADP+ | pH 8.0, 60°C, mutant enzyme A198S/S199I | Sulfurisphaera tokodaii | |
0.0222 | - |
NADP+ | mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.023 | - |
NADP+ | pH 8.0, 60°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.0231 | - |
NADP+ | wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.026 | - |
NADP+ | pH 8.0, 60°C, mutant enzyme S199I | Sulfurisphaera tokodaii | |
0.0263 | - |
NADP+ | mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.0901 | - |
NADP+ | wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.0979 | - |
NADP+ | mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.187 | - |
NADP+ | mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
17.2 | - |
NAD+ | mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
17.2 | - |
NAD+ | pH 8.0, 60°C, mutant enzyme A198S/S199I | Sulfurisphaera tokodaii | |
18.9 | - |
NAD+ | wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
19.4 | - |
NAD+ | mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
25.8 | - |
NAD+ | mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
25.8 | - |
NAD+ | pH 8.0, 60°C, mutant enzyme S199I | Sulfurisphaera tokodaii | |
26.4 | - |
NAD+ | pH 8.0, 60°C, wild-type enzyme | Sulfurisphaera tokodaii | |
26.4 | - |
NAD+ | wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
29.3 | - |
NAD+ | mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
56000 | - |
4 * 56000, SDS-PAGE | Sulfurisphaera tokodaii |
56400 | - |
4 * 56400, calculated from amino acid sequence | Sulfurisphaera tokodaii |
56400 | - |
4 * 56400, calculated from sequence | Sulfurisphaera tokodaii |
227000 | - |
gel filtration | Sulfurisphaera tokodaii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + NAD+ + H2O | Sulfurisphaera tokodaii | NAD+ is a poor co-substrate | 3-phospho-D-glycerate + NADH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | Sulfurisphaera tokodaii | - |
3-phospho-D-glycerate + NADPH + H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | Sulfurisphaera tokodaii | - |
3-phospho-D-glycerate + NADPH + 2 H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | Sulfurisphaera tokodaii 7 | - |
3-phospho-D-glycerate + NADPH + 2 H+ | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sulfurisphaera tokodaii | - |
- |
- |
Sulfurisphaera tokodaii | Q96XP0 | - |
- |
Sulfurisphaera tokodaii 7 | Q96XP0 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Sulfurisphaera tokodaii |
HisTrap column chromatography and Superdex 200 gel filtration | Sulfurisphaera tokodaii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-glyceraldehyde 3-phosphate + NAD+ + H2O | NAD+ is a poor co-substrate | Sulfurisphaera tokodaii | 3-phospho-D-glycerate + NADH + H+ | - |
? | |
D-glyceraldehyde 3-phosphate + NAD+ + H2O | the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM | Sulfurisphaera tokodaii | 3-phospho-D-glycerate + NADH + 2 H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NAD+ + H2O | the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM | Sulfurisphaera tokodaii 7 | 3-phospho-D-glycerate + NADH + 2 H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | - |
Sulfurisphaera tokodaii | 3-phospho-D-glycerate + NADPH + H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | - |
Sulfurisphaera tokodaii | 3-phospho-D-glycerate + NADPH + 2 H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM. K-type allosterism in which the positive cooperativity is abolished with concomitant activation by D-glucose 1-phosphate | Sulfurisphaera tokodaii | 3-phospho-D-glycerate + NADPH + 2 H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | - |
Sulfurisphaera tokodaii 7 | 3-phospho-D-glycerate + NADPH + 2 H+ | - |
ir | |
D-glyceraldehyde 3-phosphate + NADP+ + H2O | the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM. K-type allosterism in which the positive cooperativity is abolished with concomitant activation by D-glucose 1-phosphate | Sulfurisphaera tokodaii 7 | 3-phospho-D-glycerate + NADPH + 2 H+ | - |
ir |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 56000, SDS-PAGE | Sulfurisphaera tokodaii |
homotetramer | 4 * 56400, calculated from amino acid sequence | Sulfurisphaera tokodaii |
tetramer | 4 * 56000, SDS-PAGE | Sulfurisphaera tokodaii |
tetramer | 4 * 56400, calculated from sequence | Sulfurisphaera tokodaii |
Synonyms | Comment | Organism |
---|---|---|
GAP dehydrogenase | - |
Sulfurisphaera tokodaii |
GAPN | - |
Sulfurisphaera tokodaii |
non-phosphorylating GAP dehydrogenase | - |
Sulfurisphaera tokodaii |
St-GAPN | - |
Sulfurisphaera tokodaii |
STK_24770 | locus name | Sulfurisphaera tokodaii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
assay at | Sulfurisphaera tokodaii |
85 | - |
- |
Sulfurisphaera tokodaii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
at 60°C | Sulfurisphaera tokodaii |
8 | - |
assay at | Sulfurisphaera tokodaii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | poor cofactor | Sulfurisphaera tokodaii | |
NADP+ | preferred cofactor | Sulfurisphaera tokodaii | |
NADP+ | the enzyme prefers NADP+ to NAD+. NAD+ is a poor co-substrate | Sulfurisphaera tokodaii |
Organism | Comment | Expression |
---|---|---|
Sulfurisphaera tokodaii | ATP, ADP, AMP, D-glucose 6-phosphate and galactose 1-phosphate exhibit no effect on enzyme activity | additional information |
Sulfurisphaera tokodaii | a slight increase in enzyme activity is observed in the presence of D-fructose 6-phosphate and coenzyme A (1.1fold) | up |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.378 | - |
NAD+ | pH 8.0, 60°C, wild-type enzyme | Sulfurisphaera tokodaii | |
0.378 | - |
NAD+ | wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.434 | - |
NAD+ | mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.434 | - |
NAD+ | pH 8.0, 60°C, mutant enzyme S199I | Sulfurisphaera tokodaii | |
0.637 | - |
NAD+ | wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
0.639 | - |
NAD+ | mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
1 | - |
NAD+ | mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
1 | - |
NAD+ | pH 8.0, 60°C, mutant enzyme A198S/S199I | Sulfurisphaera tokodaii | |
1.63 | - |
NAD+ | mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
48.1 | - |
NADP+ | mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
48.1 | - |
NADP+ | pH 8.0, 60°C, mutant enzyme S199I | Sulfurisphaera tokodaii | |
61.5 | - |
NADP+ | mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
61.5 | - |
NADP+ | pH 8.0, 60°C, mutant enzyme A198S/S199I | Sulfurisphaera tokodaii | |
71.4 | - |
NADP+ | mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
96.6 | - |
NADP+ | pH 8.0, 60°C, wild-type enzyme | Sulfurisphaera tokodaii | |
96.6 | - |
NADP+ | wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
188 | - |
NADP+ | mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii | |
203 | - |
NADP+ | wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60°C | Sulfurisphaera tokodaii |