Information on EC 3.6.1.28 - thiamine-triphosphatase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.6.1.28
-
RECOMMENDED NAME
GeneOntology No.
thiamine-triphosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
thiamine triphosphate + H2O = thiamine diphosphate + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoric ester hydrolysis
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
Thiamine metabolism
-
-
thiamine triphosphate metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
thiamine-triphosphate phosphohydrolase
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CAS REGISTRY NUMBER
COMMENTARY hide
9068-47-7
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + H2O
ADP + phosphate
show the reaction diagram
CTP + H2O
CDP + phosphate
show the reaction diagram
GTP + H2O
GDP + phosphate
show the reaction diagram
ITP + H2O
IDP + phosphate
show the reaction diagram
thiamin triphosphate + H2O
thiamine diphosphate + phosphate
show the reaction diagram
-
-
-
-
?
thiamine diphosphate + H2O
thiamine monophosphate + phosphate
show the reaction diagram
thiamine triphosphate + H2O
thiamine diphosphate + phosphate
show the reaction diagram
UTP + H2O
UDP + phosphate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
thiamine triphosphate + H2O
thiamine diphosphate + phosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
-
7.3% of the activation with Mg2+
Br-
-
stimulation by anions with the following order of effectiveness: SCN-, I-, NO3-, Br-, Cl-, membrane-associated enzyme
Cl-
-
stimulation by anions with the following decreasing order of effectiveness: SCN-, I-, NO3-, Br-, Cl-, membrane-associated enzyme
I-
-
stimulation by anions with the following decreasing order of effectiveness: SCN-, I-, NO3-, Br-, Cl-, membrane-associated enzyme
Ni2+
-
30% of the activation with Mg2+
NO3-
-
stimulation by anions with the following decreasing order of effectiveness: SCN-, I-, NO3-, Br-, Cl-, membrane-associated enzyme. The effect of NO3- is maximal at pH 7-8, it is negligible at pH 6.5
SCN-
-
stimulation by anions with the following decreasing order of effectiveness: SCN-, I-, NO3-, Br-, Cl-, membrane-associated enzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
([2-[3-(4-Amino-2-methyl-pyrimidin-5-ylmethyl)-4-methyl-thiazol-5-yl]-ethoxy]-phosphonomethyl-phosphinoylmethyl)-phosphonic acid
-
;
1-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-3-(2-hydroxyethyl)-2-methylpyridinium
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soluble enzyme: 50% inhibition at 2 mM. Membrane-bound enzyme: no inhibition up to 5 mM
4,4'-diisothiocyanostilbene-2,2'-disulfonic acid
-
1.0 mM, 62% inhibition of membrane-associated enzyme
4-acetamide-4'-isothiocyanostilbene-2,2'-disulfonic acid
-
1.0 mM, 36% inhibition of membrane-associated enzyme
5,5'-dithiobis-2-nitrobenzoic acid
Br-
-
soluble enzyme
chlorpromazine
Cl-
-
soluble enzyme
Colchicine
-
1.0 mM, 16% inhibition
deoxycholate
desipramine
F-
-
5 mM, 55% inhibition
I-
-
soluble enzyme
imipramine
-
microsomal and soluble enzyme
Mg2+
-
excess of Mg2+ over thiamine triphosphate inhibits
N-dodecyl-N,N-dimethyl-3-amino-1-propanesulfonate
-
-
SCN-
-
soluble enzyme
SDS
-
IC50: about 0.3% W/v. The inhibition is partially reversible, probably due to correct refolding of the denatured enzyme
thiamine beta,gamma-methylene phosphonate
-
-
thiamine diphosphate
thiamine phosphate
-
0.75 mM, 12% inhibition
thiamine triphosphate
vanadate
-
1.0 mM, 27% inhibition of membrane-associated enzyme
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
deoxycholate
-
-
Genapol X-080
-
activates
-
Lubrol Px
-
activates
Thesit
-
activates
-
Triton X-100
-
activates
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.7
ATP
-
in presence of 8 mM ATP
0.1043
ITP
-
-
0.6
Mg2+-thiamine triphosphate complex
-
-
0.0211 - 0.126
thiamin triphosphate
0.008 - 2.8
thiamine triphosphate
additional information
additional information
-
kinetics
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.24
ATP
Homo sapiens
-
in presence of 8 mM Mg2+
0.019 - 240
thiamine triphosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
14 - 8000
thiamine triphosphate
2224
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.2
([2-[3-(4-Amino-2-methyl-pyrimidin-5-ylmethyl)-4-methyl-thiazol-5-yl]-ethoxy]-phosphonomethyl-phosphinoylmethyl)-phosphonic acid
-
-
2
1-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-3-(2-hydroxyethyl)-2-methylpyridinium
-
soluble enzyme
0.03 - 0.15
Ca2+
0.95
thiamine triphosphate
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.007 - 0.02
Zn2+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0003
quail
-
activity in quadriceps
0.0006
quail
-
activity in brain; activity in heart
0.0008
quail
-
activity in kidney
0.0024
quail
-
activity in liver
0.0114
-
activity in brain
0.0125
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activity in skeletal muscle
0.0162
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activity in kidney
0.033
-
duodenum
0.041
-
purified GST-tagged thiamine triphosphatase, 37C, pH 8.2, 4 mM MgCl2, 0.5 mM thiamine triphosphate
0.048
-
wild type enzyme from supernatant, at 37C, 50 mM Na-TAPS pH 8.5, 8 mM MgCl2, 0.5 mM thiamine triphosphate
0.052
-
activity in quadriceps
0.057
-
cerebellum
0.09
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K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 5 mM Na-EDTA
0.169
-
trapezius muscle
0.189
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activity in kidney
0.217
-
heart
0.22
-
lymph node
0.275
-
spleen
0.299
-
activity in liver
0.309
-
brain
0.333
-
lung
0.354
-
liver
0.377
-
kidney
0.437
-
activity in brain
0.45
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K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.04 mM ZnSO4
0.55
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.2 mM ZnSO4
0.59
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 4 mM CaCl2
0.63
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.1 mM ZnSO4
0.932
-
activity in pectoralis muscle
0.99
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 1 mM CaCl2
1.7
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2
1.9
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 0.1 mM ZnSO4
2.55
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K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 4 mM MgCl2 + 0.04 mM ZnSO4
3.08
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K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 1 mM MnCl2
3.3
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.05 mM MnCl2
3.45
-
K85E-mutated GST-tagged thiamine triphosphatase, pH 8.2, in the presence of 0.1 mM MnCl2
4.5
-
wild type thiamine triphosphatase from supernatant
140
-
purified recombinant enzyme
225
-
pure untagged recombinant thiamine triphosphatase, 37C, pH 8.2
648
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
-
membrane-bound enzyme
6.5 - 7
-
membrane-associated enzyme
6.8
-
wild type enzyme
7.5
-
in presence of activating anion, membrane-associated enzyme
7.8
-
and a second optimum at pH 6.5, microsomal enzyme
8.3
-
assay at
8.5 - 9
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soluble enzyme
8.7
-
-
8.9
-
-
9 - 9.5
-
soluble enzyme
9
-
GST-tagged thiamine triphosphatase
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
-
about 55% of maximal activity at pH 5.0 and pH 8.0, membrane-associated enzyme
7 - 9.5
-
pH 7.0: about 60% of maximal activity, pH 9.5: about 75% of maximal activity
7.5 - 9.5
-
pH 7.5: about 30% of maximal activity, pH 9.5: about 50% of maximal activity
8 - 10
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pH 8.0: about 60% of maximal activity, pH 10.0: about 75% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
44
-
soluble enzyme
50
-
soluble enzyme
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
metastatic prostate cancer cells
Manually annotated by BRENDA team
-
non-metastatic human colorectal adenocarcinoma cells
Manually annotated by BRENDA team
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33 nmol per min per mg of wet weight
Manually annotated by BRENDA team
-
metastatic lung cancer cells
Manually annotated by BRENDA team
-
high activity
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25550
-
gel filtration
26300
-
brain enzyme, gel filtration
27200
-
lung enzyme, gel filtration
27600
-
trapezius muscle enzyme, gel filtration
28300
-
kidney enzyme, gel filtration
28400
-
liver enzyme, gel filtratuon
29000
-
spleen enzyme, gel filtration
29700
-
liver enzyme, gel filtratuon
30000
-
enzyme from liver and brain, gel filtration
31000
-
enzyme from embryo, gel filtration
31700
-
duodenum enzyme, gel filtration
32000
-
muscle enzyme, gel filtration
32200
-
brain enzyme, gel filtration
33350
-
gel filtration
34000
-
kidney enzyme, gel filtration
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 25000, enzyme solution structure determination by NMR spectroscopy, binding of Mg2+ induces only a minor local conformational change, whereas ThTP binding causes a more global conformational change, structural model for the mThTPase-ThTP-Mg2+ ternary complex, free mThTPase has an open cleft fold, the enzyme in the ternary complex adopts a tunnel fold, PDB ID 2JMU, overview
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
complex with tripolyphosphate, to 2.3 A resolution and docking solution of substrate thiamine triphosphate. The thiazole ring of the thiamine forms a stacking interaction with Trp53, and the aminopyrimidine ring lies in a pocket defined by His76, Pro191 and Ile195. The triphosphate moiety occupies a similar position as in the enzyme-tripolyphosphate complex, with interactions involving the side chains of Lys11, Arg55, Arg57, Lys65, Arg125 and Lys193
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
-
GST-tagged wild type enzyme activity was nearly zero at pH 5.0
35
-
at pH 5.0: 50% loss of activity after 83 min, at pH 7.6: about 50% loss of activity after 10 min
55 - 60
-
30 min, stable
68
-
30 min, 50% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
not inactivated by freezing and thawing
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.05 M Tris-HCl buffer, stable for several months
-
0C, stable for many hours
-
4C, stable for about 2 weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
MagneGST Protein Purification System, only mutated enzymes expressed as GST fusion proteins
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli strain BL21
expression analysis
expression in Escherichia coli, the recombinant enzyme is completely functional
-
overexpression in Escherichia coli as a glutathione S-transferase fusion protein
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E63Q
-
reduced activity
E78K
-
reduced activity
E85K
-
reduced activity
D147A
-
about 30% of wild-type catalytic efficiency
D37A
-
about 0.5% of wild-type catalytic efficiency
E81A
-
about 30% of wild-type catalytic efficiency
K11A
-
about 15% of wild-type catalytic efficiency
K193A
-
about 60% of wild-type catalytic efficiency
K65A
-
more than 1000fold decrease in kcat value
W53A
-
2.5fold increase in catalytic efficiency
Y39A
-
about 30% of wild-type catalytic efficiency
Y79A
-
about 5% of wild-type catalytic efficiency
K85E
-
decreased solubility, reduced catalytic activity
additional information