Cloned (Comment) | Organism |
---|---|
Escherichia coli strain KL21 (DE3)pLysS is transformed with the expression vector coding for a histidine-tagged wild-type or mutant hTPK1 | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D100N | mutation reduces turnover-number markedly | Homo sapiens |
D133N | mutation causes a selective decrease in the ratio of turnover-number to Km-value for thiamine | Homo sapiens |
D71N | mutation reduces turnover-number markedly | Homo sapiens |
D73N | mutation reduces turnover-number markedly | Homo sapiens |
Q96E | mutation causes an 2.5fold increase in the ratio of turnover-number to Km-value for thiamine compared to the wild-type | Homo sapiens |
R131G | mutation decreases the ratio of turnover-number to Km-value for ATP | Homo sapiens |
S74A | mutation causes a 1.4fold increase in turnover number, the Km-value for ATP is 2fold that of the wild-type enzyme | Homo sapiens |
T99A | mutation decreases the ratio of turnover-number to Km-value for ATP | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.039 | - |
thiamine | pH 7.5, 37°C, mutant enzyme Q96E | Homo sapiens | |
0.076 | - |
thiamine | pH 7.5, 37°C, mutant enzyme R131G | Homo sapiens | |
0.16 | - |
thiamine | pH 7.5, 37°C, mutant enzyme T99A | Homo sapiens | |
0.21 | - |
thiamine | pH 7.5, 37°C, wild-type enzyme | Homo sapiens | |
0.23 | - |
thiamine | pH 7.5, 37°C, mutant enzyme S74A | Homo sapiens | |
0.65 | - |
ATP | pH 7.5, 37°C, mutant enzyme D133N | Homo sapiens | |
0.82 | - |
thiamine | pH 7.5, 37°C, mutant enzyme D133N | Homo sapiens | |
1 | - |
ATP | pH 7.5, 37°C, mutant enzyme Q96E | Homo sapiens | |
1.2 | - |
ATP | pH 7.5, 37°C, wild-type enzyme | Homo sapiens | |
2 | - |
ATP | pH 7.5, 37°C, mutant enzyme R131G | Homo sapiens | |
2.4 | - |
ATP | pH 7.5, 37°C, mutant enzyme S74A | Homo sapiens | |
3.5 | - |
ATP | pH 7.5, 37°C, mutant enzyme T99A | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | divalent cation required, 7.3% of the activation with Mg2+ | Homo sapiens | |
Co2+ | divalent cation required, 90.6% of the activation with Mg2+ | Homo sapiens | |
Mg2+ | divalent cation required, best activation with Mg2+ | Homo sapiens | |
Mn2+ | divalent cation required, 16.8% of the activation with Mg2+ | Homo sapiens | |
Zn2+ | divalent cation required, 27% of the activation with Mg2+ | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
30000 | - |
2 * 30000, SDS-PAGE | Homo sapiens |
61000 | - |
gel filtration | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9H3S4 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + thiamine = AMP + thiamine diphosphate | ping pong mechanism | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0602 | - |
- |
Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + thiamine | - |
Homo sapiens | AMP + thiamine diphosphate | - |
? | |
CTP + thiamine | 31.4% of the activity with ATP | Homo sapiens | CMP + thiamine diphosphate | - |
? | |
GTP + thiamine | 18.3% of the activity with ATP | Homo sapiens | GMP + thiamine diphosphate | - |
? | |
UTP + thiamine | 207% of the activity with ATP | Homo sapiens | UMP + thiamine diphosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 30000, SDS-PAGE | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0002 | - |
ATP | pH 7.5, 37°C, mutant enzyme D100N | Homo sapiens | |
0.0002 | - |
thiamine | pH 7.5, 37°C, mutant enzyme D100N | Homo sapiens | |
0.0003 | - |
ATP | pH 7.5, 37°C, mutant enzyme D73N | Homo sapiens | |
0.0003 | - |
thiamine | pH 7.5, 37°C, mutant enzyme D73N | Homo sapiens | |
0.0005 | - |
ATP | pH 7.5, 37°C, mutant enzyme D71N | Homo sapiens | |
0.0005 | - |
thiamine | pH 7.5, 37°C, mutant enzyme D71N | Homo sapiens | |
0.026 | - |
ATP | pH 7.5, 37°C, mutant enzyme T99A | Homo sapiens | |
0.026 | - |
thiamine | pH 7.5, 37°C, mutant enzyme T99A | Homo sapiens | |
0.028 | - |
ATP | pH 7.5, 37°C, mutant enzyme R131G | Homo sapiens | |
0.028 | - |
thiamine | pH 7.5, 37°C, mutant enzyme R131G | Homo sapiens | |
0.034 | - |
ATP | pH 7.5, 37°C, mutant enzyme D133N | Homo sapiens | |
0.034 | - |
thiamine | pH 7.5, 37°C, mutant enzyme D133N | Homo sapiens | |
0.045 | - |
ATP | pH 7.5, 37°C, mutant enzyme Q96E | Homo sapiens | |
0.045 | - |
thiamin | pH 7.5, 37°C, mutant enzyme Q96E | Homo sapiens | |
0.069 | - |
ATP | pH 7.5, 37°C, wild-type enzyme | Homo sapiens | |
0.069 | - |
thiamine | pH 7.5, 37°C, wild-type enzyme | Homo sapiens | |
0.094 | - |
ATP | pH 7.5, 37°C, mutant enzyme S74A | Homo sapiens | |
0.094 | - |
thiamine | pH 7.5, 37°C, mutant enzyme S74A | Homo sapiens |