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Literature summary extracted from

  • Ranjbarian, F.; Vodnala, M.; Vodnala, S.M.; Rofougaran, R.; Thelander, L.; Hofer, A.
    Trypanosoma brucei thymidine kinase is tandem protein consisting of two homologous parts, which together enable efficient substrate binding (2012), J. Biol. Chem., 287, 17628-17636.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment 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 Cloned (Comment) Organism
2.7.1.21 expressed in Escherichia coli BL21-CodonPlus (DE3) cells Trypanosoma brucei

Inhibitors

EC Number Inhibitors Comment 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 Comment 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 UniProt Comment Textmining
2.7.1.21 Trypanosoma brucei
-
-
-
2.7.1.21 Trypanosoma brucei TC221
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.21 Ni-NTA agarose column chromatography Trypanosoma brucei

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.21 ATP + 5-fluorodeoxyuridine
-
 Trypanosoma brucei ADP + 5-fluorodeoxyuridine 5'-phosphate
-
 ?
2.7.1.21 ATP + deoxyadenosine
-
 Trypanosoma brucei ADP + dAMP
-
 ?
2.7.1.21 ATP + deoxyadenosine
-
 Trypanosoma brucei TC221 ADP + dAMP
-
 ?
2.7.1.21 ATP + deoxycytidine
-
 Trypanosoma brucei ADP + dCMP
-
 ?
2.7.1.21 ATP + deoxycytidine
-
 Trypanosoma brucei TC221 ADP + dCMP
-
 ?
2.7.1.21 ATP + deoxyguanosine about 20% activity compared to thymidine Trypanosoma brucei ADP + dGMP
-
 ?
2.7.1.21 ATP + deoxyguanosine about 20% activity compared to thymidine Trypanosoma brucei TC221 ADP + dGMP
-
 ?
2.7.1.21 ATP + deoxyinosine about 30% activity compared to thymidine Trypanosoma brucei ADP + dIMP
-
 ?
2.7.1.21 ATP + deoxyinosine about 30% activity compared to thymidine Trypanosoma brucei TC221 ADP + dIMP
-
 ?
2.7.1.21 ATP + deoxyuridine about 185% activity compared to thymidine Trypanosoma brucei ADP + dUMP
-
 ?
2.7.1.21 ATP + thymidine best substrate combination, 100% activity Trypanosoma brucei ADP + dTMP
-
 ?
2.7.1.21 ATP + uridine
-
 Trypanosoma brucei ADP + UMP
-
 ?
2.7.1.21 CTP + thymidine
-
 Trypanosoma brucei CDP + dTMP
-
 ?
2.7.1.21 GTP + thymidine
-
 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 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 Trypanosoma brucei TC221 ?
-
 ?
2.7.1.21 UTP + thymidine
-
 Trypanosoma brucei UDP + dTMP
-
 ?

Subunits

EC Number Subunits Comment 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 Comment 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
-
 thymidine 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 3.5
-
 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 Comment 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 Comment 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