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Literature summary extracted from

  • Szyniarowski Piot, S.P.; Lakaye Bernar, L.B.; Czerniecki Ja, C.J.; Makarchikov Alexander , M.A.; Wins Pierr, W.P.; Margineanu Ilc, M.I.; Coumans Bernar, C.B.; Grisar Thierr, G.T.; Bettendorff Lucie, B.L.
    Pig tissues express a catalytically inefficient 25-kDa thiamine triphosphatase: insight in the catalytic mechanisms of this enzyme (2005), Biochim. Biophys. Acta, 1725, 93-102.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.6.1.28 expressed in Escherichia coli strain BL21 Sus scrofa
3.6.1.28 expressed in Escherichia coli strain BL21 Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
3.6.1.28 E63Q reduced activity Homo sapiens
3.6.1.28 E78K reduced activity Homo sapiens
3.6.1.28 E85K reduced activity Homo sapiens
3.6.1.28 K85E decreased solubility, reduced catalytic activity Sus scrofa

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.6.1.28 Ca2+
-
 Homo sapiens
3.6.1.28 Zn2+ pH 8.5, IC50: 0.007 mM Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.6.1.28 0.15
-
 thiamine triphosphate wild type enzyme Homo sapiens
3.6.1.28 0.2
-
 thiamine triphosphate
-
 Sus scrofa
3.6.1.28 0.25
-
 thiamine triphosphate GST-fused E63Q mutant enzyme Homo sapiens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.6.1.28 Mg2+ 5 mM, in the absence of Mg2+ the activity is still 40-50% of the maximum Sus scrofa
3.6.1.28 Mg2+ enzyme is 5% active in the absence of Mg2+ Homo sapiens
3.6.1.28 Mn2+ 0.1 mM Homo sapiens
3.6.1.28 Zn2+ 0.01-0.05 mM at pH 8.0 activates GST-tagged thiamine triphosphatase by about 30% in the presence of Mg2+ Sus scrofa
3.6.1.28 Zn2+ in the presence of Mg2+ at pH 8 Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.6.1.28 25000
-
 SDS-PAGE Sus scrofa
3.6.1.28 25000
-
 SDS-PAGE Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
3.6.1.28 Homo sapiens Q9BU02
-
-
3.6.1.28 Sus scrofa
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.6.1.28 MagneGST Protein Purification System, only mutated enzymes expressed as GST fusion proteins Sus scrofa
3.6.1.28 MagneGST Protein Purification System, only mutated enzymes expressed as GST fusion proteins Homo sapiens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.6.1.28 brain
-
 Sus scrofa
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.6.1.28 0.041
-
 purified GST-tagged thiamine triphosphatase, 37°C, pH 8.2, 4 mM MgCl2, 0.5 mM thiamine triphosphate Sus scrofa
3.6.1.28 0.048
-
 wild type enzyme from supernatant, at 37°C, 50 mM Na-TAPS pH 8.5, 8 mM MgCl2, 0.5 mM thiamine triphosphate Sus scrofa
3.6.1.28 0.09
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 5 mM Na-EDTA Sus scrofa
3.6.1.28 0.45
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.04 mM ZnSO4 Sus scrofa
3.6.1.28 0.55
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.2 mM ZnSO4 Sus scrofa
3.6.1.28 0.59
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 4 mM CaCl2 Sus scrofa
3.6.1.28 0.63
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.1 mM ZnSO4 Sus scrofa
3.6.1.28 0.99
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 1 mM CaCl2 Sus scrofa
3.6.1.28 1.7
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 Sus scrofa
3.6.1.28 1.9
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 0.1 mM ZnSO4 Sus scrofa
3.6.1.28 2.55
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 0.04 mM ZnSO4 Sus scrofa
3.6.1.28 3.08
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 1 mM MnCl2 Sus scrofa
3.6.1.28 3.3
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.05 mM MnCl2 Sus scrofa
3.6.1.28 3.45
-
 K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.1 mM MnCl2 Sus scrofa
3.6.1.28 4.5
-
 wild type thiamine triphosphatase from supernatant Homo sapiens
3.6.1.28 225
-
 pure untagged recombinant thiamine triphosphatase, 37°C, pH 8.2 Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.6.1.28 thiamine triphosphate + H2O
-
 Sus scrofa thiamine diphosphate + phosphate
-
 ?
3.6.1.28 thiamine triphosphate + H2O
-
 Homo sapiens thiamine diphosphate + phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.6.1.28 25-kDa thiamine triphosphatase
-
 Sus scrofa
3.6.1.28 25-kDa thiamine triphosphatase
-
 Homo sapiens
3.6.1.28 ThTPase
-
 Sus scrofa
3.6.1.28 ThTPase
-
 Homo sapiens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.6.1.28 5
-
 GST-tagged wild type enzyme activity was nearly zero at pH 5.0 Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.6.1.28 6.8
-
 wild type enzyme Sus scrofa
3.6.1.28 8.5
-
 wild type enzyme Homo sapiens
3.6.1.28 9
-
 GST-tagged thiamine triphosphatase Sus scrofa
3.6.1.28 9.4
-
 GST-fused E63Q mutant enzyme Homo sapiens
3.6.1.28 9.4
-
 K85E-mutated thiamine triphosphatase fused to GST Sus scrofa

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.6.1.28 0.03
-
 Ca2+ GST-fused E78K mutant enzyme Homo sapiens
3.6.1.28 0.15
-
 Ca2+ GST-fused wild type enzyme Homo sapiens

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.6.1.28 0.007
-
 pH 8.5, IC50: 0.007 mM Homo sapiens Zn2+