Information on EC 2.7.6.2 - thiamine diphosphokinase

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

EC NUMBER
COMMENTARY
2.7.6.2
-
RECOMMENDED NAME
GeneOntology No.
thiamine diphosphokinase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + thiamine = AMP + thiamine diphosphate
show the reaction diagram
ping pong mechanism
-
ATP + thiamine = AMP + thiamine diphosphate
show the reaction diagram
partial ping pong mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
diphosphate transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Metabolic pathways
-
-
thiamin diphosphate biosynthesis III (Staphylococcus)
-
-
thiamin diphosphate biosynthesis IV (eukaryotes)
-
-
thiamin salvage III
-
-
Thiamine metabolism
-
-
vitamin B1 metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:thiamine diphosphotransferase
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
At1g02880 (AtTPK1)
-
-
At2g44750 (AtTPK2)
F4IV16
-
ATP:thiamin pyrophosphotransferase
-
-
-
-
hTPK1
-
-
-
-
Plasmodium falciparum thiamine pyrophosphokinase
-
-
pyrophosphokinase, thiamin
-
-
-
-
TDPK
-
-
thiamin kinase
-
-
-
-
thiamin pyrophosphokinase
-
-
-
-
thiamin pyrophosphokinase
F4IV16
-
thiamin pyrophosphokinase
-
-
thiamin pyrophosphokinase
I3PU26
-
thiamin pyrophosphokinase
-
-
thiamin pyrophosphotransferase
-
-
-
-
thiamin:ATP pyrophosphotransferase
-
-
-
-
thiamine diphosphokinase
-
-
thiamine kinase
-
-
-
-
thiamine pyrophokinase
-
-
thiamine pyrophosphokinase
-
encoded by tpk-1
thiamine pyrophosphokinase
-
-
thiamine pyrophosphokinase
-
-
thiamine pyrophosphokinase
-
-
thiamine pyrophosphokinase
Q59N99
-
thiamine pyrophosphokinase
-
-
thiamine pyrophosphokinase
-
-
thiaminokinase
-
-
-
-
TPK
-
-
-
-
TPK
Q59N99
-
Tpk1
-
-
TPTase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9026-24-8
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
P41888
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
Q9H3S4
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
Q59N99
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
I3PU26
-
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
P41888
the enzyme is a regulator of thiamine metabolism, phosphate metabolism, mating, and growth
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
the enzyme is important for the formation of the coenzyme thiamine diphosphate
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
P30636
the tpk-1 gene functions cell nonautonomously, as the expression of wild-type tpk-1 in one tissue can residue the function of other tissues that express only mutant tpk-1. Thiamine diphosphate can be transported across the cell membrane
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
enzyme is involved in biosynthesis of thiamine diphosphate
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
importance of TPP/thiamine in the alpha-oxidation pathway. Most TPP is tightly bound to peroxisomal proteins, most likely to 2-hydroxyacyl-CoA lyase 1, HACL1, or membranes
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
Q59N99
importance of TPP/thiamine in the alpha-oxidation pathway. Most TPP is tightly bound to peroxisomal proteins, most likely to 2-hydroxyacyl-CoA lyase 1, HACL1, or membranes
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
F4IV16
thiamine pyrophosphokinase is required for thiamine cofactor activation in Arabidopsis
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
F4IV16
no TMP kinase activity is shown by either isozymes TPK1 or TPK2
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
Paracoccus denitrificans 12442
-
-
-
-
?
CTP + thiamine
CMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
CTP + thiamine
CMP + thiamine diphosphate
show the reaction diagram
Q9H3S4
31.4% of the activity with ATP
-
-
?
GTP + thiamine
GMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
GTP + thiamine
GMP + thiamine diphosphate
show the reaction diagram
Q9H3S4
18.3% of the activity with ATP
-
-
?
oxythiamine + ATP
oxythiamine diphosphate + AMP
show the reaction diagram
-
-
-
-
?
pyrithiamine + ATP
pyrithiamine diphosphate + AMP
show the reaction diagram
-
inhibits the substrate thiamine
-
-
?
pyrithiamine + ATP
pyrithiamine diphosphate + AMP
show the reaction diagram
-
its specific activity is 30% lower in comparison to its natural substrate thiamine
-
-
?
thiamin + ATP
thiamindiphosphate + AMP
show the reaction diagram
F4IV16
-
-
-
?
thiamine + ATP
thiamine diphosphate + AMP
show the reaction diagram
-
-
-
-
?
thiamine + ATP
thiamine diphosphate + AMP
show the reaction diagram
-
-
-
-
-
thiamine + ATP
thiamine diphosphate + AMP
show the reaction diagram
-
TPP plays a critical role in the carbohydrate and energy metabolism. It functions as a prosthetic group for the mitochondrial enzyme complexes like pyruvate dehydrogenase, alpha-ketoglutarate dehydrogenase and branched chain alpha-keto acid dehydrogenase. In addition, TPP is involved in the cytosolic pentose pathway functioning as coenzyme for transketolase.
-
-
?
UTP + thiamine
UMP + thiamine diphosphate
show the reaction diagram
-
-
-
-
?
UTP + thiamine
UMP + thiamine diphosphate
show the reaction diagram
Q9H3S4
207% of the activity with ATP
-
-
?
GTP + thiamine
GMP + thiamine diphosphate
show the reaction diagram
-
GTP is the preferred nucleotide
-
-
-
additional information
?
-
-
the YcfN protein acts as a nonspecific adenosylcobinamide kinase enzyme, which is not associated with adenosylcobalamine biosynthesis, but it allows a CobU-deficient strain of serovar Typhimurium to synthesize enough adenosylcobalamine from cobinamide so that the cell can grow. YcfN is required for the phosphorylation of thiamine in the thiamine salvaging pathway of Escherichia coli
-
-
-
additional information
?
-
-
three common intronic variants in the maternal and fetal thiamine pyrophosphokinase gene (TPK1) are associated with birth weight, overview
-
-
-
additional information
?
-
-
thiamine binding site structure, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
P41888
the enzyme is a regulator of thiamine metabolism, phosphate metabolism, mating, and growth
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
the enzyme is important for the formation of the coenzyme thiamine diphosphate
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
P30636
the tpk-1 gene functions cell nonautonomously, as the expression of wild-type tpk-1 in one tissue can residue the function of other tissues that express only mutant tpk-1. Thiamine diphosphate can be transported across the cell membrane
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
-
importance of TPP/thiamine in the alpha-oxidation pathway. Most TPP is tightly bound to peroxisomal proteins, most likely to 2-hydroxyacyl-CoA lyase 1, HACL1, or membranes
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
Q59N99
importance of TPP/thiamine in the alpha-oxidation pathway. Most TPP is tightly bound to peroxisomal proteins, most likely to 2-hydroxyacyl-CoA lyase 1, HACL1, or membranes
-
-
?
thiamin + ATP
thiamindiphosphate + AMP
show the reaction diagram
F4IV16
-
-
-
?
thiamine + ATP
thiamine diphosphate + AMP
show the reaction diagram
-
-
-
-
?
thiamine + ATP
thiamine diphosphate + AMP
show the reaction diagram
-
-
-
-
-
thiamine + ATP
thiamine diphosphate + AMP
show the reaction diagram
-
TPP plays a critical role in the carbohydrate and energy metabolism. It functions as a prosthetic group for the mitochondrial enzyme complexes like pyruvate dehydrogenase, alpha-ketoglutarate dehydrogenase and branched chain alpha-keto acid dehydrogenase. In addition, TPP is involved in the cytosolic pentose pathway functioning as coenzyme for transketolase.
-
-
?
ATP + thiamine
AMP + thiamine diphosphate
show the reaction diagram
F4IV16
thiamine pyrophosphokinase is required for thiamine cofactor activation in Arabidopsis
-
-
?
additional information
?
-
-
the YcfN protein acts as a nonspecific adenosylcobinamide kinase enzyme, which is not associated with adenosylcobalamine biosynthesis, but it allows a CobU-deficient strain of serovar Typhimurium to synthesize enough adenosylcobalamine from cobinamide so that the cell can grow. YcfN is required for the phosphorylation of thiamine in the thiamine salvaging pathway of Escherichia coli
-
-
-
additional information
?
-
-
three common intronic variants in the maternal and fetal thiamine pyrophosphokinase gene (TPK1) are associated with birth weight, overview
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ATP
Q59N99
-
myo-Inositol 1-diphosphate
-
coenzyme
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
-
divalent cation required, 7.3% of the activation with Mg2+
Co2+
-
divalent cation required, 90.6% of the activation with Mg2+
Co2+
-
absolute requirement for a divalent cation such as Mg2+, Mn2+ or Co2+. In presence of 2 mM ATP maximal activity occurs with 1 mM Co2+
Mg2+
-
required, maximal activation at 20 mM
Mg2+
-
divalent cation required, best activation with Mg2+
Mg2+
-
absolute requirement for a divalent cation such as Mg2+, Mn2+ or Co2+. In presence of 2 mM ATP maximal activity occurs with above 3 mM Mg2+
Mg2+
-
required
Mg2+
-
for optimal activity the ratio of magnesium to ATP must be fixed at 0.6. The Mg(ATP)26- species might by the actual diphosphoryl donor
Mg2+
P41888
Km: 3 mM
Mg2+
-
Optimal activity obtained by addition of 2-4 mM Mg2+.
Mg2+
-
-
Mg2+
-
ligands electronic density map and Mg2+ coordination structure, the delta-oxygen from D113 and D115 and one oxygen atom from each phosphate group of the thiamine-PNP. The last coordination of this Mg2+ involves an oxygen atom from the free inorganic phosphate, overview
Mn2+
-
activates at low concentrations even more than Mg2+
Mn2+
-
divalent cation required, 16.8% of the activation with Mg2+
Mn2+
-
absolute requirement for a divalent cation such as Mg2+, Mn2+ or Co2+. In presence of 2 mM ATP maximal activity occurs with 0.5 mM Mn2+
Zn2+
-
divalent cation required, 27% of the activation with Mg2+
Zn2+
-
Substitution of magnesium by Zn2+ decreases the enzyme activity by 85%.
Mn2+
-
Substitution of magnesium by Mn2+ decreases the enzyme activity by 42%.
additional information
-
ligand binding structures, overview
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ALT-711
-
low-affinity inhibitor, mixed-mode inhibition with a major role for competitive inhibition
-
AMP
-
0.5 mM, 8% inhibition
Butylthiamine
-
competitive with respect to thiamine
Chloroethylthiamine
-
weak
diphosphate
-
competitive with respect to thiamine
diphosphate
-
-
diphosphate
-
10 mM, complete inhibition
diphosphate
-
0.5 mM, only 3% inhibition
ethylthiamine
-
competitive with respect to thiamine
iodoacetamide
-
dithiothreitol prevents inhibition
iodoacetamide
-
10 mM
iodoacetamide
-
3.3 mM, 23.3% inhibition, activity is recovered to approximately 90% on incubation with a 10fold excess of dithiothreitol
KCN
-
little effect
molybdate
-
little effect
NaF
-
little effect
NEM
-
dithiothreitol prevents inhibition
Oxythiamine
-
weak
Oxythiamine
-
competitive with respect to thiamine
Oxythiamine
-
0.5 mM; 4% inhibition
PCMB
-
dithiothreitol prevents inhibition
PCMB
-
0.1 mM, complete inhibition, completely restored by 10 mM dithiothreitol
PCMB
-
1 mM, 55.1% inhibition, activity is recovered to approximately 90% on incubation with a 10fold excess of dithiothreitol
PCMB
-
0.5 mM, 36% inhibition
pyrithiamine
-
-
pyrithiamine
-
competitive with respect to thiamine
pyrithiamine
-
0.5 mM, 77% inhibition
pyrithiamine
-
-
thiamine diphosphate
-
0.5 mM; product feedback inhibition
thiamine diphosphate
-
-
thiamine diphosphate
-
0.5 mM; 95% inhibition
Thiamine monophosphate
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
myo-Inositol
-
stimulates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0041
ATP
-
pH 7.8, 37C
0.059
ATP
-
pH 7.8, 37C
0.38
ATP
-
pH 9.0, 37C
0.65
ATP
-
pH 7.5, 37C, mutant enzyme D133N
1
ATP
-
pH 7.5, 37C, mutant enzyme Q96E
1.2
ATP
-
pH 7.5, 37C, wild-type enzyme
1.9
ATP
P41888
pH 8.6, 37C
2
ATP
-
pH 7.5, 37C, mutant enzyme R131G
2.4
ATP
-
pH 7.5, 37C, mutant enzyme S74A
3.5
ATP
-
pH 7.5, 37C, mutant enzyme T99A
18.3
ATP
-
pH 8.1, 37C
7.6
CTP
-
pH 8.1, 37C
0.021
GTP
-
-
2.6
GTP
-
pH 8.1, 37C
0.062
pyrithiamine
-
-
0.00015
thiamine
-
-
0.00096
thiamine
F4IV16
AtTPK1; pH 8.0, 37C, isozyme TPK1
0.00128
thiamine
F4IV16
AtTPK2; pH 8.0, 37C, isozyme TPK2
0.00464
thiamine
-
pH 8.1, 37C
0.006
thiamine
P41888
pH 8.6, 37C
0.01
thiamine
-
pH 9.0, 37C
0.039
thiamine
-
pH 7.5, 37C, mutant enzyme Q96E
0.073
thiamine
-
thiamine monophosphate is not accepted
0.076
thiamine
-
pH 7.5, 37C, mutant enzyme R131G
0.16
thiamine
-
pH 7.5, 37C, mutant enzyme T99A
0.21
thiamine
-
pH 7.5, 37C, wild-type enzyme
0.23
thiamine
-
pH 7.5, 37C, mutant enzyme S74A
0.82
thiamine
-
pH 7.5, 37C, mutant enzyme D133N
2.9
thiamine
-
-
11.2
UTP
-
pH 8.1, 37C
4.5
MgATP2-
-
-
additional information
additional information
-
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0002
ATP
-
pH 7.5, 37C, mutant enzyme D100N
0.0003
ATP
-
pH 7.5, 37C, mutant enzyme D73N
0.0005
ATP
-
pH 7.5, 37C, mutant enzyme D71N
0.026
ATP
-
pH 7.5, 37C, mutant enzyme T99A
0.028
ATP
-
pH 7.5, 37C, mutant enzyme R131G
0.034
ATP
-
pH 7.5, 37C, mutant enzyme D133N
0.045
ATP
-
pH 7.5, 37C, mutant enzyme Q96E
0.069
ATP
-
pH 7.5, 37C, wild-type enzyme
0.094
ATP
-
pH 7.5, 37C, mutant enzyme S74A
0.045
thiamin
-
pH 7.5, 37C, mutant enzyme Q96E
0.0002
thiamine
-
pH 7.5, 37C, mutant enzyme D100N
0.0003
thiamine
-
pH 7.5, 37C, mutant enzyme D73N
0.0005
thiamine
-
pH 7.5, 37C, mutant enzyme D71N
0.026
thiamine
-
pH 7.5, 37C, mutant enzyme T99A
0.028
thiamine
-
pH 7.5, 37C, mutant enzyme R131G
0.034
thiamine
-
pH 7.5, 37C, mutant enzyme D133N
0.069
thiamine
-
pH 7.5, 37C, wild-type enzyme
0.094
thiamine
-
pH 7.5, 37C, mutant enzyme S74A
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.8
diphosphate
-
-
0.019
pyrithiamine
-
-
0.2
Thiamine monophosphate
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.000061
-
-
0.000367
-
recombinant enzyme
0.00165
-
-
0.009
-
PfTPK3, with 4 mM magnesium ions in the enzyme assay
0.017
-
PfTPK2, with 4 mM magnesium ions in the enzyme assay; substrate: pyrithiamine, PfTPK
0.027
-
PfTPK, with 4 mM magnesium ions in the enzyme assay
0.0602
-
-
999
-
no activity with thiochrome
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7
F4IV16
isozymes TPK1 or TPK2
7.1 - 7.3
-
-
8 - 9
-
in Tris buffer
8.3 - 9.3
-
in 0.028 M phosphate/glycylglycine buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 8
F4IV16
;
6.4 - 8
-
pH 6.4: about 40% of maximal activity, pH 8.0: about 50% of maximal activity
6.8 - 9.3
-
pH 6.8: about 65% of maximal activity, pH 8.3-9.3: optimum
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
F4IV16
AtTPK2; isozyme TPK2
45
F4IV16
AtTPK1; isozyme TPK1
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37 - 50
F4IV16
37C: optimal activity of TPK2, 50C: TPK2 is completely inactive
45 - 55
F4IV16
TPK1
50
F4IV16
AtTPK2 is completely inactive
55
F4IV16
AtTPK1 is still active
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.2
-
isoelectric focusing
4.5
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
low expression
Manually annotated by BRENDA team
-
highly expressed
Manually annotated by BRENDA team
-
low expression
Manually annotated by BRENDA team
-
highly expressed
Manually annotated by BRENDA team
-
low expression
Manually annotated by BRENDA team
-
highly expressed
Manually annotated by BRENDA team
-
low expression
Manually annotated by BRENDA team
additional information
F4IV16
isozymes TPK1 and TPK2 are biochemically redundant cytosolic proteins that are similarly expressed throughout different plant tissues
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Q59N99
exclusively
Manually annotated by BRENDA team
-
Peroxisomes are devoid of thiamine pyrophosphokinase activity.
Manually annotated by BRENDA team
F4IV16
isozymes TPK1 and TPK2
Manually annotated by BRENDA team
additional information
-
no enzyme localization in the peroxisome, subcellular localization analysis, overview
-
Manually annotated by BRENDA team
additional information
Q59N99
no enzyme localization in the peroxisome, subcellular localization analysis, overview
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Bacillus subtilis (strain 168)
Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
Candida albicans (strain SC5314 / ATCC MYA-2876)
Candida albicans (strain SC5314 / ATCC MYA-2876)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30000
-
gel filtration
642745, 642748
38000
-
for PfTPK3, SDS-PAGE analysis followed by Western blotting
672476
43000
-
for PfTPK2, SDS-PAGE analysis followed by Western blotting
672476
44000
-
gel filtration
642749
47000
-
for PfTPK, SDS-PAGE analysis followed by Western blotting
672476
49000
-
gel filtration
642753
61000
-
gel filtration
642743
96000
-
aggregate of the functional dimer, equilibrium sedimentation
642749
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
I3PU26
x * 28000-30000, SDS-PAGE
dimer
-
2 * 30000, SDS-PAGE
dimer
-
2 * 230000, SDS-PAGE
dimer
-
gel filtration using a calibrated Superdex S-200 column
dimer
-
three-dimensional enzyme structure, modelling, overview
dimer
Paracoccus denitrificans 12442
-
2 * 230000, SDS-PAGE
-
homodimer
-
-
homotetramer
-
x-ray crystallography
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme in complex with ATP analogue AMP-PNP and thiamine, hanging drop vapor diffusion method, mixing of 500 nl of protein solution with 500 nl of reservoir solution containing 17.5-20% PEG 4000, 0.2 M MgCl2 , 0.1 M Tris, and 20% glycerol, pH 7.0-pH 7.5, and equilibration against 1 ml or reservoir solution, X-ray diffraction structure determination and analysis at 1.96-2.1 A resolution
-
complexed with thiamin diphosphate-Mg2+ or thiamin-Mg2+, hanging drop vapor diffusion method, using 1.15-1.3 M sodium citrate, 0.1 M Na-HEPES, pH 8.6, at 22C
-
Crystallization experiments were set up by incubating mouse TPK with pyrithiamine and MgATP2- prior to adding the equilibrium mixture of enzyme, substrate, and products to the previously determined crystallization conditions.
-
recombinant enzyme, hanging drop vapor diffusion method
-
hanging-drop vapor diffusion method, crystals of the complex of native recombinant enzyme with thiamine
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
F4IV16
inactivation of isozyme TPK2
676560
55
F4IV16
isozyme TPK1 shows remaining activity
676560
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme becomes increasingly unstable with progressive purification
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant CA1462 from Escherichia coli
-
partial
-
nickel-chelate chromatography
-
partial
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
isozymes TPK1 and TPK2, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression in Escherichia coli strain BL21, expression of fluorescent tagged isozymes in cytosol of mesophyll protoplasts
F4IV16
gene CA1462, expression in Escherichia coli
-
Escherichia coli strain KL21 (DE3)pLysS is transformed with the expression vector coding for a histidine-tagged wild-type or mutant hTPK1
-
TPK1 gene, located in 7q34-36, genotyping in 964 individuals from 220 families of European ancestry revealing an association between maternal rs228584 genotype and offspring birth weight
-
expressed in Escherichia coli BL21 cells
-
overexpression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
during embryogenesis, total thiamin diphosphokinase transcripts increase abruptly in early development, and decrease to about half of the peak shortly after hatching
I3PU26
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D100N
-
mutation reduces turnover-number markedly
D133N
-
mutation causes a selective decrease in the ratio of turnover-number to Km-value for thiamine
D71N
-
mutation reduces turnover-number markedly
Q96E
-
mutation causes an 2.5fold increase in the ratio of turnover-number to Km-value for thiamine compared to the wild-type
R131G
-
mutation decreases the ratio of turnover-number to Km-value for ATP
S74A
-
mutation causes a 1.4fold increase in turnover number, the Km-value for ATP is 2fold that of the wild-type enzyme
T99A
-
mutation decreases the ratio of turnover-number to Km-value for ATP
additional information
F4IV16
construction of a double knockout mutant lacking isozymes TPK1 and TPK2, the mutant shows severely depressed levels of TPP and elevated levels of free thiamine compared to the wild-type enzyme
D73N
-
mutation reduces turnover-number markedly
additional information
-
three common intronic variants in the maternal and fetal thiamine pyrophosphokinase gene (TPK1) are associated with birth weight, overview
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
-
Malaria is a devastating and quite often deadly parasitic disease, resulting in significant public health problems in the tropics. The inevitable emergence of drug-resistant forms requires the continuous discovery and development of new antimalarials. These drugs should only target the parasite, with only harmless or no effects on the human host. Thus, ideal drug targets would be peculiarities in the parasite metabolism, such as vitamin B1 and B6 biosyntheses that are absent in its host.
additional information
-
Administration of oxythiamine to animals produces primarily metabolic aberrations such as lethargy and anorexia while the administration of pyrithiamine to animals produces neurological problems similar to Wernicke-Korsakoff Syndrome.