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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.6.2 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

EC Number Protein Variants Comment Organism
2.7.6.2 D100N mutation reduces turnover-number markedly Homo sapiens
2.7.6.2 D133N mutation causes a selective decrease in the ratio of turnover-number to Km-value for thiamine Homo sapiens
2.7.6.2 D71N mutation reduces turnover-number markedly Homo sapiens
2.7.6.2 D73N mutation reduces turnover-number markedly Homo sapiens
2.7.6.2 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
2.7.6.2 R131G mutation decreases the ratio of turnover-number to Km-value for ATP Homo sapiens
2.7.6.2 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
2.7.6.2 T99A mutation decreases the ratio of turnover-number to Km-value for ATP Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.6.2 0.039
-
 thiamine pH 7.5, 37°C, mutant enzyme Q96E Homo sapiens
2.7.6.2 0.076
-
 thiamine pH 7.5, 37°C, mutant enzyme R131G Homo sapiens
2.7.6.2 0.16
-
 thiamine pH 7.5, 37°C, mutant enzyme T99A Homo sapiens
2.7.6.2 0.21
-
 thiamine pH 7.5, 37°C, wild-type enzyme Homo sapiens
2.7.6.2 0.23
-
 thiamine pH 7.5, 37°C, mutant enzyme S74A Homo sapiens
2.7.6.2 0.65
-
 ATP pH 7.5, 37°C, mutant enzyme D133N Homo sapiens
2.7.6.2 0.82
-
 thiamine pH 7.5, 37°C, mutant enzyme D133N Homo sapiens
2.7.6.2 1
-
 ATP pH 7.5, 37°C, mutant enzyme Q96E Homo sapiens
2.7.6.2 1.2
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
2.7.6.2 2
-
 ATP pH 7.5, 37°C, mutant enzyme R131G Homo sapiens
2.7.6.2 2.4
-
 ATP pH 7.5, 37°C, mutant enzyme S74A Homo sapiens
2.7.6.2 3.5
-
 ATP pH 7.5, 37°C, mutant enzyme T99A Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.6.2 Ca2+ divalent cation required, 7.3% of the activation with Mg2+ Homo sapiens
2.7.6.2 Co2+ divalent cation required, 90.6% of the activation with Mg2+ Homo sapiens
2.7.6.2 Mg2+ divalent cation required, best activation with Mg2+ Homo sapiens
2.7.6.2 Mn2+ divalent cation required, 16.8% of the activation with Mg2+ Homo sapiens
2.7.6.2 Zn2+ divalent cation required, 27% of the activation with Mg2+ Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.6.2 30000
-
 2 * 30000, SDS-PAGE Homo sapiens
2.7.6.2 61000
-
 gel filtration Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
2.7.6.2 Homo sapiens Q9H3S4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.6.2
-
 Homo sapiens

Reaction

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.6.2 0.0602
-
-
 Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.6.2 ATP + thiamine
-
 Homo sapiens AMP + thiamine diphosphate
-
 ?
2.7.6.2 CTP + thiamine 31.4% of the activity with ATP Homo sapiens CMP + thiamine diphosphate
-
 ?
2.7.6.2 GTP + thiamine 18.3% of the activity with ATP Homo sapiens GMP + thiamine diphosphate
-
 ?
2.7.6.2 UTP + thiamine 207% of the activity with ATP Homo sapiens UMP + thiamine diphosphate
-
 ?

Subunits

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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.6.2 0.0002
-
 ATP pH 7.5, 37°C, mutant enzyme D100N Homo sapiens
2.7.6.2 0.0002
-
 thiamine pH 7.5, 37°C, mutant enzyme D100N Homo sapiens
2.7.6.2 0.0003
-
 ATP pH 7.5, 37°C, mutant enzyme D73N Homo sapiens
2.7.6.2 0.0003
-
 thiamine pH 7.5, 37°C, mutant enzyme D73N Homo sapiens
2.7.6.2 0.0005
-
 ATP pH 7.5, 37°C, mutant enzyme D71N Homo sapiens
2.7.6.2 0.0005
-
 thiamine pH 7.5, 37°C, mutant enzyme D71N Homo sapiens
2.7.6.2 0.026
-
 ATP pH 7.5, 37°C, mutant enzyme T99A Homo sapiens
2.7.6.2 0.026
-
 thiamine pH 7.5, 37°C, mutant enzyme T99A Homo sapiens
2.7.6.2 0.028
-
 ATP pH 7.5, 37°C, mutant enzyme R131G Homo sapiens
2.7.6.2 0.028
-
 thiamine pH 7.5, 37°C, mutant enzyme R131G Homo sapiens
2.7.6.2 0.034
-
 ATP pH 7.5, 37°C, mutant enzyme D133N Homo sapiens
2.7.6.2 0.034
-
 thiamine pH 7.5, 37°C, mutant enzyme D133N Homo sapiens
2.7.6.2 0.045
-
 ATP pH 7.5, 37°C, mutant enzyme Q96E Homo sapiens
2.7.6.2 0.045
-
 thiamin pH 7.5, 37°C, mutant enzyme Q96E Homo sapiens
2.7.6.2 0.069
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
2.7.6.2 0.069
-
 thiamine pH 7.5, 37°C, wild-type enzyme Homo sapiens
2.7.6.2 0.094
-
 ATP pH 7.5, 37°C, mutant enzyme S74A Homo sapiens
2.7.6.2 0.094
-
 thiamine pH 7.5, 37°C, mutant enzyme S74A Homo sapiens