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Literature summary for 3.6.1.28 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
Search for more literature for 3.6.1.28

Cloned(Commentary)

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

Protein Variants

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

Inhibitors

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

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ 5 mM, in the absence of Mg2+ the activity is still 40-50% of the maximum Sus scrofa
Mg2+ enzyme is 5% active in the absence of Mg2+ Homo sapiens
Mn2+ 0.1 mM Homo sapiens
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
Zn2+ in the presence of Mg2+ at pH 8 Homo sapiens

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Homo sapiens Q9BU02
-
-
Sus scrofa
-
-
-

Purification (Commentary)

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

Source Tissue

Source Tissue Comment Organism Textmining
brain
-
 Sus scrofa
-

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Synonyms

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

Temperature Stability [°C]

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

pH Optimum

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

Ki Value [mM]

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

IC50 Value

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