BRENDA - Enzyme Database show

Trypanosoma brucei thymidine kinase is tandem protein consisting of two homologous parts, which together enable efficient substrate binding

Ranjbarian, F.; Vodnala, M.; Vodnala, S.M.; Rofougaran, R.; Thelander, L.; Hofer, A.; J. Biol. Chem. 287, 17628-17636 (2012)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.21
dithiothreitol
the enzyme is stimulated by dithiothreitol up to a concentration of 2 mM
Trypanosoma brucei
2.7.1.21
KCl
weak stimulation
Trypanosoma brucei
2.7.1.21
NaCl
weak stimulation
Trypanosoma brucei
2.7.1.21
potassium acetate
weak stimulation
Trypanosoma brucei
Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.21
expressed in Escherichia coli BL21-CodonPlus (DE3) cells
Trypanosoma brucei
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.21
sulfate
competitive inhibitor of ATP binding
Trypanosoma brucei
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.0078
-
thymidine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.03
-
5-fluorodeoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.046
-
thymidine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.08
-
deoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.098
-
ATP
thymidine kinase domain 2, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.18
-
ATP
full-length enzyme, with 0.0005 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.21
-
ATP
full-length enzyme, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.22
-
ATP
full-length enzyme, with 0.005 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.32
-
ATP
full-length enzyme, with 0.5 mM deoxyinosine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.43
-
deoxyuridine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
2.5
-
deoxyinosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
2.6
-
deoxyguanosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
7.8
-
uridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
8.4
-
deoxyadenosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.21
Trypanosoma brucei
-
-
-
2.7.1.21
Trypanosoma brucei TC221
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.21
Ni-NTA agarose column chromatography
Trypanosoma brucei
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 + 5-fluorodeoxyuridine
-
722758
Trypanosoma brucei
ADP + 5-fluorodeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + deoxyadenosine
-
722758
Trypanosoma brucei
ADP + dAMP
-
-
-
?
2.7.1.21
ATP + deoxyadenosine
-
722758
Trypanosoma brucei TC221
ADP + dAMP
-
-
-
?
2.7.1.21
ATP + deoxycytidine
-
722758
Trypanosoma brucei
ADP + dCMP
-
-
-
?
2.7.1.21
ATP + deoxycytidine
-
722758
Trypanosoma brucei TC221
ADP + dCMP
-
-
-
?
2.7.1.21
ATP + deoxyguanosine
about 20% activity compared to thymidine
722758
Trypanosoma brucei
ADP + dGMP
-
-
-
?
2.7.1.21
ATP + deoxyguanosine
about 20% activity compared to thymidine
722758
Trypanosoma brucei TC221
ADP + dGMP
-
-
-
?
2.7.1.21
ATP + deoxyinosine
about 30% activity compared to thymidine
722758
Trypanosoma brucei
ADP + dIMP
-
-
-
?
2.7.1.21
ATP + deoxyinosine
about 30% activity compared to thymidine
722758
Trypanosoma brucei TC221
ADP + dIMP
-
-
-
?
2.7.1.21
ATP + deoxyuridine
about 185% activity compared to thymidine
722758
Trypanosoma brucei
ADP + dUMP
-
-
-
?
2.7.1.21
ATP + thymidine
best substrate combination, 100% activity
722758
Trypanosoma brucei
ADP + dTMP
-
-
-
?
2.7.1.21
ATP + uridine
-
722758
Trypanosoma brucei
ADP + UMP
-
-
-
?
2.7.1.21
CTP + thymidine
-
722758
Trypanosoma brucei
CDP + dTMP
-
-
-
?
2.7.1.21
GTP + thymidine
-
722758
Trypanosoma brucei
GDP + dTMP
-
-
-
?
2.7.1.21
additional information
domain 1 of thymidine kinase is catalytically inactive but improves substrate binding by domain 2. The enzyme shows the following order of catalytic efficiencies: thymidine > deoxyuridine >> deoxyinosine > deoxyguanosine. The enzyme shows no activity with deoxyadenosine and deoxycytidine
722758
Trypanosoma brucei
?
-
-
-
-
2.7.1.21
additional information
domain 1 of thymidine kinase is catalytically inactive but improves substrate binding by domain 2. The enzyme shows the following order of catalytic efficiencies: thymidine > deoxyuridine >> deoxyinosine > deoxyguanosine. The enzyme shows no activity with deoxyadenosine and deoxycytidine
722758
Trypanosoma brucei TC221
?
-
-
-
-
2.7.1.21
UTP + thymidine
-
722758
Trypanosoma brucei
UDP + dTMP
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.7.1.21
monomer
-
Trypanosoma brucei
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.047
-
deoxyadenosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.34
-
deoxyguanosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.8
-
deoxyinosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
1.5
-
uridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.1
-
ATP
full-length enzyme, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.1
-
thymidine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.5
-
thymidine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.6
-
5-fluorodeoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.9
-
ATP
thymidine kinase domain 2, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
5.1
-
deoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
5.3
-
deoxyuridine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.21
7.8
-
-
Trypanosoma brucei
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.21
dithiothreitol
the enzyme is stimulated by dithiothreitol up to a concentration of 2 mM
Trypanosoma brucei
2.7.1.21
KCl
weak stimulation
Trypanosoma brucei
2.7.1.21
NaCl
weak stimulation
Trypanosoma brucei
2.7.1.21
potassium acetate
weak stimulation
Trypanosoma brucei
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
expressed in Escherichia coli BL21-CodonPlus (DE3) cells
Trypanosoma brucei
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.21
sulfate
competitive inhibitor of ATP binding
Trypanosoma brucei
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.0078
-
thymidine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.03
-
5-fluorodeoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.046
-
thymidine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.08
-
deoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.098
-
ATP
thymidine kinase domain 2, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.18
-
ATP
full-length enzyme, with 0.0005 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.21
-
ATP
full-length enzyme, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.22
-
ATP
full-length enzyme, with 0.005 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.32
-
ATP
full-length enzyme, with 0.5 mM deoxyinosine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.43
-
deoxyuridine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
2.5
-
deoxyinosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
2.6
-
deoxyguanosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
7.8
-
uridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
8.4
-
deoxyadenosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.21
Ni-NTA agarose column chromatography
Trypanosoma brucei
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 + 5-fluorodeoxyuridine
-
722758
Trypanosoma brucei
ADP + 5-fluorodeoxyuridine 5'-phosphate
-
-
-
?
2.7.1.21
ATP + deoxyadenosine
-
722758
Trypanosoma brucei
ADP + dAMP
-
-
-
?
2.7.1.21
ATP + deoxyadenosine
-
722758
Trypanosoma brucei TC221
ADP + dAMP
-
-
-
?
2.7.1.21
ATP + deoxycytidine
-
722758
Trypanosoma brucei
ADP + dCMP
-
-
-
?
2.7.1.21
ATP + deoxycytidine
-
722758
Trypanosoma brucei TC221
ADP + dCMP
-
-
-
?
2.7.1.21
ATP + deoxyguanosine
about 20% activity compared to thymidine
722758
Trypanosoma brucei
ADP + dGMP
-
-
-
?
2.7.1.21
ATP + deoxyguanosine
about 20% activity compared to thymidine
722758
Trypanosoma brucei TC221
ADP + dGMP
-
-
-
?
2.7.1.21
ATP + deoxyinosine
about 30% activity compared to thymidine
722758
Trypanosoma brucei
ADP + dIMP
-
-
-
?
2.7.1.21
ATP + deoxyinosine
about 30% activity compared to thymidine
722758
Trypanosoma brucei TC221
ADP + dIMP
-
-
-
?
2.7.1.21
ATP + deoxyuridine
about 185% activity compared to thymidine
722758
Trypanosoma brucei
ADP + dUMP
-
-
-
?
2.7.1.21
ATP + thymidine
best substrate combination, 100% activity
722758
Trypanosoma brucei
ADP + dTMP
-
-
-
?
2.7.1.21
ATP + uridine
-
722758
Trypanosoma brucei
ADP + UMP
-
-
-
?
2.7.1.21
CTP + thymidine
-
722758
Trypanosoma brucei
CDP + dTMP
-
-
-
?
2.7.1.21
GTP + thymidine
-
722758
Trypanosoma brucei
GDP + dTMP
-
-
-
?
2.7.1.21
additional information
domain 1 of thymidine kinase is catalytically inactive but improves substrate binding by domain 2. The enzyme shows the following order of catalytic efficiencies: thymidine > deoxyuridine >> deoxyinosine > deoxyguanosine. The enzyme shows no activity with deoxyadenosine and deoxycytidine
722758
Trypanosoma brucei
?
-
-
-
-
2.7.1.21
additional information
domain 1 of thymidine kinase is catalytically inactive but improves substrate binding by domain 2. The enzyme shows the following order of catalytic efficiencies: thymidine > deoxyuridine >> deoxyinosine > deoxyguanosine. The enzyme shows no activity with deoxyadenosine and deoxycytidine
722758
Trypanosoma brucei TC221
?
-
-
-
-
2.7.1.21
UTP + thymidine
-
722758
Trypanosoma brucei
UDP + dTMP
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.1.21
monomer
-
Trypanosoma brucei
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.047
-
deoxyadenosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.34
-
deoxyguanosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.8
-
deoxyinosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
1.5
-
uridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.1
-
ATP
full-length enzyme, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.1
-
thymidine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.5
-
thymidine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.6
-
5-fluorodeoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
3.9
-
ATP
thymidine kinase domain 2, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
5.1
-
deoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
5.3
-
deoxyuridine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.21
7.8
-
-
Trypanosoma brucei
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.0056
-
deoxyadenosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.13
-
deoxyguanosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.19
-
uridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.32
-
deoxyinosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
12
-
deoxyuridine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
15
-
ATP
full-length enzyme, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
40
-
ATP
thymidine kinase domain 2, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
63
-
deoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
76
-
thymidine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
120
-
5-fluorodeoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
400
-
thymidine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.1.21
0.0056
-
deoxyadenosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.13
-
deoxyguanosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.19
-
uridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
0.32
-
deoxyinosine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
12
-
deoxyuridine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
15
-
ATP
full-length enzyme, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
40
-
ATP
thymidine kinase domain 2, with 0.05 mM thymidine as cosubstrate, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
63
-
deoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
76
-
thymidine
thymidine kinase domain 2, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
120
-
5-fluorodeoxyuridine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei
2.7.1.21
400
-
thymidine
full-length enzyme, at pH 7.8 and 37°C
Trypanosoma brucei