BRENDA - Enzyme Database show

Tight binding of deoxyribonucleotide triphosphates to human thymidine kinase 2 expressed in Escherichia coli. Purification and partial characterization of its dimeric and tetrameric forms

Barroso, J.F.; Elholm, M.; Flatmark, T.; Biochemistry 42, 15158-15169 (2003)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.21
DNA and amino acid sequence determination, expression in Escherichia coli as maltose binding fusion protein in inclusion bodies, coexpression of GroEL/ES molecular chaperone system improves enzyme solubility only marginally, but ethanol supplementation contributes to enzyme recovery due to activation of heat shock response
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
additional information
-
additional information
kinetics
Homo sapiens
2.7.1.21
0.0047
-
ATP
dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0109
-
ATP
tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.018
-
2'-deoxycytidine
tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.025
-
2'-deoxycytidine
dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.05
-
2'-deoxythymidine
tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.054
-
2'-deoxythymidine
dimeric enzyme, pH 7.5, 25°C
Homo sapiens
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.1.21
cytosol
-
Homo sapiens
5829
-
2.7.1.21
mitochondrion
-
Homo sapiens
5739
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.21
Mg2+
-
Homo sapiens
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.21
additional information
-
oligomeric states have MWs of 152200 and 95900, as determined by gel filtration
Homo sapiens
2.7.1.21
48800
-
recombinant enzyme, gel filtration
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
Homo sapiens
-
ADP + 2'-deoxycytidine 5'-phosphate
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
Homo sapiens
-
ADP + 2'-deoxythymidine 5'-phosphate
-
-
ir
2.7.1.21
ATP + 2'-deoxyuridine
Homo sapiens
-
ADP + 2'-deoxyuridine 5'-phosphate
-
-
ir
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.21
Homo sapiens
O00142
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.21
recombinant enzyme as maltose binding fusion protein from Escherichia coli
Homo sapiens
Reaction
EC Number
Reaction
Commentary
Organism
2.7.1.21
ATP + thymidine = ADP + dTMP
molecular docking of substrates and inhibitors, active site structure modeling
Homo sapiens
Renatured (Commentary)
EC Number
Commentary
Organism
2.7.1.21
coexpression of GroEL/ES molecular chaperone system improves recombinant enzyme solubility from inclusion bodies only marginally, but ethanol supplementation contributes to enzyme recovery due to activation of heat shock response
Homo sapiens
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
2.7.1.21
additional information
expression in nonproliferating tissue
Homo sapiens
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
-
661066
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
-
661066
Homo sapiens
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxyuridine
-
661066
Homo sapiens
ADP + 2'-deoxyuridine 5'-phosphate
-
-
-
ir
Subunits
EC Number
Subunits
Commentary
Organism
2.7.1.21
dimer
enzyme appears as dimer and tetramer
Homo sapiens
2.7.1.21
More
structure modeling
Homo sapiens
2.7.1.21
tetramer
enzyme appears as dimer and tetramer
Homo sapiens
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.21
37
-
-
Homo sapiens
Temperature Range [°C]
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
2.7.1.21
25
37
highest activity within this temperature range at 37°C
Homo sapiens
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.21
7.5
-
assay at
Homo sapiens
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.21
ATP
-
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
DNA and amino acid sequence determination, expression in Escherichia coli as maltose binding fusion protein in inclusion bodies, coexpression of GroEL/ES molecular chaperone system improves enzyme solubility only marginally, but ethanol supplementation contributes to enzyme recovery due to activation of heat shock response
Homo sapiens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.21
ATP
-
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
additional information
-
additional information
kinetics
Homo sapiens
2.7.1.21
0.0047
-
ATP
dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.0109
-
ATP
tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.018
-
2'-deoxycytidine
tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.025
-
2'-deoxycytidine
dimeric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.05
-
2'-deoxythymidine
tetrameric enzyme, pH 7.5, 25°C
Homo sapiens
2.7.1.21
0.054
-
2'-deoxythymidine
dimeric enzyme, pH 7.5, 25°C
Homo sapiens
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.1.21
cytosol
-
Homo sapiens
5829
-
2.7.1.21
mitochondrion
-
Homo sapiens
5739
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.21
Mg2+
-
Homo sapiens
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.21
additional information
-
oligomeric states have MWs of 152200 and 95900, as determined by gel filtration
Homo sapiens
2.7.1.21
48800
-
recombinant enzyme, gel filtration
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
Homo sapiens
-
ADP + 2'-deoxycytidine 5'-phosphate
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
Homo sapiens
-
ADP + 2'-deoxythymidine 5'-phosphate
-
-
ir
2.7.1.21
ATP + 2'-deoxyuridine
Homo sapiens
-
ADP + 2'-deoxyuridine 5'-phosphate
-
-
ir
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
recombinant enzyme as maltose binding fusion protein from Escherichia coli
Homo sapiens
Renatured (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
coexpression of GroEL/ES molecular chaperone system improves recombinant enzyme solubility from inclusion bodies only marginally, but ethanol supplementation contributes to enzyme recovery due to activation of heat shock response
Homo sapiens
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
2.7.1.21
additional information
expression in nonproliferating tissue
Homo sapiens
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.21
ATP + 2'-deoxycytidine
-
661066
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxythymidine
-
661066
Homo sapiens
ADP + 2'-deoxythymidine 5'-phosphate
-
-
-
ir
2.7.1.21
ATP + 2'-deoxyuridine
-
661066
Homo sapiens
ADP + 2'-deoxyuridine 5'-phosphate
-
-
-
ir
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.1.21
dimer
enzyme appears as dimer and tetramer
Homo sapiens
2.7.1.21
More
structure modeling
Homo sapiens
2.7.1.21
tetramer
enzyme appears as dimer and tetramer
Homo sapiens
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.21
37
-
-
Homo sapiens
Temperature Range [°C] (protein specific)
EC Number
Temperature Minimum [°C]
Temperature Maximum [°C]
Commentary
Organism
2.7.1.21
25
37
highest activity within this temperature range at 37°C
Homo sapiens
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.21
7.5
-
assay at
Homo sapiens