Information on EC 2.7.1.50 - hydroxyethylthiazole kinase

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

EC NUMBER
COMMENTARY
2.7.1.50
-
RECOMMENDED NAME
GeneOntology No.
hydroxyethylthiazole kinase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole = ADP + 4-methyl-5-(2-phosphonooxyethyl)thiazole
show the reaction diagram
-
-
-
-
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole = ADP + 4-methyl-5-(2-phosphonooxyethyl)thiazole
show the reaction diagram
catalytic mechanism of phosphoryl transfer reaction and role of active site residues, e.g. R121 and C198, calculated on the basis of data on crystal structures of ribokinase-like kinases, transition state stabilization involves an oxyanion hole and a magnesium ion
-
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole = ADP + 4-methyl-5-(2-phosphonooxyethyl)thiazole
show the reaction diagram
SN2-like mechanism, with gamma-phosphate acting as an alcohol-activation base. Mg2+ ion 2, residues E126 and C198 play a major catalytic role, whereas Mg2+ ion 1 and C198 are responsible for product release
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
phospho group transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
4-methyl-5(beta-hydroxyethyl)thiazole salvage (yeast)
-
Metabolic pathways
-
thiamin formation from pyrithiamine and oxythiamine (yeast)
-
thiamin salvage II
-
Thiamine metabolism
-
SYSTEMATIC NAME
IUBMB Comments
ATP:4-methyl-5-(2-hydroxyethyl)thiazole 2-phosphotransferase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4-methyl-5-(beta-hydroxyethyl) thiazole kinase
-
-
4-methyl-5-(beta-hydroxyethyl)thiazole kinase
-
-
-
-
4-methyl-5-beta-hydroxyethylthiazole kinase
-
-
4-methyl-5-beta-hydroxyethylthiazole kinase
-
-
4-methyl-5-beta-hydroxyethylthiazole kinase
O58877
-
4-methyl-5-beta-hydroxyethylthiazole kinase
-
-
-
-
4-methyl-5-hydroxyethylthiazole kinase
Q6FV03
C-terminal domain of TH16
hydroxyethylthiazole kinase/thiamine-phosphate pyrophosphorylase
-
bifunctional enzyme with hydroxyethylthiazole kinase and thiamine-phosphate pyrophosphorylase activity
TH kinase
-
-
-
-
THI6
Q6FV03
-
ThiK
-
-
-
-
ThiM
Q6FV03
-
THZ kinase
-
-
-
-
THZ kinase
-
-
kinase, hydroxyethylthiazole (phosphorylating)
-
-
-
-
additional information
-
the enzyme belongs to the family of ribokinase-like carbohydrate kinases
CAS REGISTRY NUMBER
COMMENTARY
9026-56-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
-
Q6FV03
UniProt
Manually annotated by BRENDA team
bv. viciae strain 3841
-
-
Manually annotated by BRENDA team
bifunctional enzyme with hydroxyethylthiazole kinase and thiamine-phosphate pyrophosphorylase activity; mutant resistant to 2-amino-4-methyl-5-beta-hydroxyethylthiazole, an antimetabolite of 4-methyl-5-beta-hydroxyethylthiazole, deficient in activity of both EC 2.5.1.3 and EC 2.7.1.50
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
-
three-quarters of the residues involved in interfacial regions are conserved, also the amino-acid residues in the nucleotide-binding, magnesium ion-binding and substrate-binding sites are conserved. The overall structure of PhThiK is similar to those of Bacillus subtilis ThiK (BsThiK) and Enterococcus faecalis V583 ThiK (EfThiK)
evolution
-, Q6FV03
THI6 is a bifunctional enzyme found in the thiamin biosynthetic pathway in eukaryotes. In prokaryotes, thiamin phosphate synthase and 4-methyl-5-hydroxyethylthiazole kinase are separate gene products
physiological function
-, Q6FV03
the N-terminal domain of THI6 catalyzes the ligation of the thiamin thiazole and pyrimidine moieties to form thiamin phosphate, and the C-terminal domain catalyzes the phosphorylation of 4-methyl-5-hydroxyethylthiazole in a salvage pathway
metabolism
-, Q6FV03
THI6 is a bifunctional enzyme of the thiamin biosynthetic pathway
additional information
-
the enzyme topology shows the typical ribokinase fold of an alpha/beta protein. Binding of the nucleotide and substrate to the ThiK enzyme do not influence the trimeric quaternary association
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ADP + 4-methyl-5-(2-hydroxyethyl)thiazole
AMP + thiazole monophosphate
show the reaction diagram
-
3.4% of the activity with ATP
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-
-
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-
-
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-, P39593
phosphate transfer occurs by an inline mechanism
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-
the bifunctioinal enzyme hydroxyethylthiazole kinase/thiamine-phosphate pyrophosphorylase catalyzes two sequential steps in the synthesis of thiamin monophosphate from hydroxyethylthiazole
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-
enzyme involved in biosynthesis of thiamine
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-, P39593
the enzyme is a salvage enzyme in the thiamin biosynthetic pathway and enables the cell to use recycled 4-methyl-5-beta-hydroxyethylthiazole as an alternative to its synthesis from 1-deoxy-D-xylulose-5-phosphate, cysteine, and tyrosine
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + 4-methyl-5-(2-phosphoethyl)thiazole
show the reaction diagram
-
-
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + 4-methyl-5-(2-phosphonooxyethyl)thiazole
show the reaction diagram
-, Q6FV03
-
-
-
?
CTP + 4-methyl-5-(2-hydroxyethyl)thiazole
CDP + thiazole monophosphate
show the reaction diagram
-
13.7% of the activity with ATP
-
-
?
dATP + 4-methyl-5-(2-hydroxyethyl)thiazole
dADP + thiazole monophosphate
show the reaction diagram
-
24.8% of the activity with ATP
-
-
?
UTP + 4-methyl-5-(2-hydroxyethyl)thiazole
UDP + thiazole monophosphate
show the reaction diagram
-
75.1% of the activity with ATP
-
-
?
GTP + 4-methyl-5-(2-hydroxyethyl)thiazole
GDP + thiazole monophosphate
show the reaction diagram
-
3.2% of the activity with ATP
-
-
?
additional information
?
-
-
ATP-dependent phosphorylation of small molecules containing hydroxymethyl groups, reaction pathway mapping, molecular mechanism optimization and quantum mechanis calculations on the basis of enzyme crystal structure, PDB code 1C3Q and 1ESQ, overview
-
-
-
additional information
?
-
-
the active site is located at the interface between adjacent subunits. Each active site contains a thiazole-binding site and a nucleotide-binding site, structure, overview. Asp residue as a catalytic base
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-
the bifunctioinal enzyme hydroxyethylthiazole kinase/thiamine-phosphate pyrophosphorylase catalyzes two sequential steps in the synthesis of thiamin monophosphate from hydroxyethylthiazole
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-
enzyme involved in biosynthesis of thiamine
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + thiazole monophosphate
show the reaction diagram
-, P39593
the enzyme is a salvage enzyme in the thiamin biosynthetic pathway and enables the cell to use recycled 4-methyl-5-beta-hydroxyethylthiazole as an alternative to its synthesis from 1-deoxy-D-xylulose-5-phosphate, cysteine, and tyrosine
-
-
?
ATP + 4-methyl-5-(2-hydroxyethyl)thiazole
ADP + 4-methyl-5-(2-phosphonooxyethyl)thiazole
show the reaction diagram
-, Q6FV03
-
-
-
?
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
10 mM, stimulates as effectively as Mg2+
Mg2+
-
10 mM, stimulates as effectively as Co2+
Mg2+
-
stabilizes the transition state and the phosphoryl goup
Mg2+
O58877, -
amino-acid residues in the magnesium ion-binding site are conserved
Mg2+
-, Q6FV03
required, two ions per enzyme molecule, binding structure involving C467, D340, and E372, overview
Mn2+
-
stimulates, 42.7% as effectively as Mg2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-Aminohydroxyethylthiazole
-
-
PCMB
-
0.01 mM, 98.5% inhibition, inhibition prevented by addition of 2-mercaptoethanol
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.79
-
ATP
-
pH 7.5, 37C
0.0057
-
hydroxyethylthiazole
-
pH 7.5, 37C
additional information
-
additional information
-
modeling of stationary points along the reaction pathway, determination of relative energies of the transition state and product
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0021
-
2-Aminohydroxyethylthiazole
-
pH 7.5, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.4
9.2
-
pH 7.4: about 80% of activity maximum, pH 9.2: about 25% of activity maximum
PDB
SCOP
CATH
ORGANISM
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Enterococcus faecalis (strain ATCC 700802 / V583)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
470000
-
-
bifunctional enzyme with hydroxyethylthiazole kinase and thiamine-phosphate pyrophosphorylase activity, gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
homohexamer
-, Q6FV03
the six protomers form a cage-like structure. Each protomer is composed of two domains, which are structurally homologous to their monofunctional bacterial counterparts. Two loop regions not found in the bacterial enzymes provide interactions between the two domains
octamer
-
8 * 60000, bifunctional enzyme with hydroxyethylthiazole kinase and thiamine-phosphate pyrophosphorylase activity, SDS-PAGE
trimer
-
residues involved in hydrogen bonds between subunits in the enzyme trimer, overview
homotrimer
O58877, -
3 * 28950 Da, crystal structure, calculated
additional information
-
as determined from crystallization data the enzyme is a trimer of identical subunits, the active site is formed at the interface between two subunits within the trimer
additional information
-
monomer structure, docking of ternary complexes, tertiary structure, molecular modelling, overview
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystallization by vapor diffusion equilibration, crystal structure at 1.5 A resolution
-
purified recombinant detagged enzyme, in complex with beta,gamma-methyleneadenosine 5'-diphosphate, thiamin phosphate, 4-amino-5-hydroxymethyl-2-trifluoromethylpyrimidine diphosphate, or 4-methyl-5-hydroxyethylthiazole phosphate, hanging-drop vapor diffusion method, mixing of0.001 ml protein solution with 0.001 ml of reservoir solution containing 0.1 M HEPES, pH 7.5, 0.2-0.25 M MgCl2, and 25-32% PEG400, 22C, 2-3 days, method optimization, X-ray diffraction structure determination and analysis at 2.6-3.3 A resolution, modeling
-, Q6FV03
enzyme with a phosphate ion occupying the position of the beta-phosphate of the nucleotide, mixing of 0.001 ml of protein solution containing 1.95 mg/ml protein in 20 mM Tris-HCl containing 200 mM NaCl and 1 mM DTT, pH 8.0, with 0.001 ml of reservoir solution containing 0.1 M CHESS buffer, pH 9.3, and 0.88 M sodium citrate as the precipitant, 20C, 1 week, 25% v/v glycerol for cryoprotection, X-ray diffraction structure determination and analysis at 1.85 A resolution
-
precipitant: sodium citrate, pH 9.3, 293 K
O58877, -
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80C, about 50% loss of activity after 1 month
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, cleavage of the tag by TEV protease, and another step of nickel affinity chromatography
-, Q6FV03
recombinant enzyme from Escherichia coli strain BL21-CodonPlus(DE3)-RIL by gel filtration, anion exchange chromatography to over 95% purity
-
heat treatment (363 K, 13 min), centrifugation, ion exchange chromatography, hydroxyapatite chromatography, gel filtration,
O58877, -
bifunctional enzyme with hydroxyethylthiazole kinase and thiamine-phosphate pyrophosphorylase activity
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
-, Q6FV03
expression in Escherichia coli strain BL21-CodonPlus(DE3)-RIL
-
expressed in Escherichia coli BL21-CodonPlus(DE3)-RIL
O58877, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C198D
-
increase in catalytic activity, catalytic field predictions