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Literature summary for 2.7.6.2 extracted from

  • Onozuka, M.; Nosaka, K.
    Steady-state kinetics and mutational studies of recombinant human thiamin pyrophosphokinase (2003), J. Nutr. Sci. Vitaminol., 49, 156-162.
    View publication on PubMed
Search for more literature for 2.7.6.2

Cloned(Commentary)

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

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 [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

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 [Da] Molecular Weight Maximum [Da] Comment Organism
30000
-
 2 * 30000, SDS-PAGE Homo sapiens
61000
-
 gel filtration Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens Q9H3S4
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
ATP + thiamine = AMP + thiamine diphosphate ping pong mechanism Homo sapiens
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0602
-
-
 Homo sapiens

Substrates and Products (Substrate)

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

Subunits Comment Organism
dimer 2 * 30000, SDS-PAGE Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0002
-
 thiamine pH 7.5, 37°C, mutant enzyme D100N Homo sapiens
0.0002
-
 ATP pH 7.5, 37°C, mutant enzyme D100N Homo sapiens
0.0003
-
 thiamine pH 7.5, 37°C, mutant enzyme D73N Homo sapiens
0.0003
-
 ATP pH 7.5, 37°C, mutant enzyme D73N Homo sapiens
0.0005
-
 thiamine pH 7.5, 37°C, mutant enzyme D71N Homo sapiens
0.0005
-
 ATP pH 7.5, 37°C, mutant enzyme D71N Homo sapiens
0.026
-
 thiamine pH 7.5, 37°C, mutant enzyme T99A Homo sapiens
0.026
-
 ATP pH 7.5, 37°C, mutant enzyme T99A Homo sapiens
0.028
-
 thiamine pH 7.5, 37°C, mutant enzyme R131G Homo sapiens
0.028
-
 ATP pH 7.5, 37°C, mutant enzyme R131G Homo sapiens
0.034
-
 thiamine pH 7.5, 37°C, mutant enzyme D133N Homo sapiens
0.034
-
 ATP pH 7.5, 37°C, mutant enzyme D133N Homo sapiens
0.045
-
 thiamin pH 7.5, 37°C, mutant enzyme Q96E Homo sapiens
0.045
-
 ATP pH 7.5, 37°C, mutant enzyme Q96E Homo sapiens
0.069
-
 thiamine pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.069
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.094
-
 thiamine pH 7.5, 37°C, mutant enzyme S74A Homo sapiens
0.094
-
 ATP pH 7.5, 37°C, mutant enzyme S74A Homo sapiens