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Literature summary extracted from

  • Wu, C.C.; Hsu, T.Y.; Chen, J.Y.
    Characterization of three essential residues in the conserved ATP-binding region of Epstein-Barr virus thymidine kinase (2005), Biochemistry, 44, 4785-4793.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.7.1.21 medicine enzyme is a target for antiviral therapy and treatment of EBV-associated malignancies Human gammaherpesvirus 4

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.21 expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Human gammaherpesvirus 4

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.21 G294A site-directed mutagenesis, about 80% reduced activity compared to the wild-type enzyme, no activity with TTP as cofactor Human gammaherpesvirus 4
2.7.1.21 G294V site-directed mutagenesis, nearly inactive mutant, no ATP binding Human gammaherpesvirus 4
2.7.1.21 K297E site-directed mutagenesis, nearly inactive mutant, no ATP binding Human gammaherpesvirus 4
2.7.1.21 K297Q site-directed mutagenesis, nearly inactive mutant, no ATP binding Human gammaherpesvirus 4
2.7.1.21 K297R site-directed mutagenesis, about 90% reduced activity compared to the wild-type enzyme, prefers GTP as cofactor, no activity with TTP as cofactor Human gammaherpesvirus 4
2.7.1.21 T298A site-directed mutagenesis, about 90% reduced activity compared to the wild-type enzyme Human gammaherpesvirus 4
2.7.1.21 T298S site-directed mutagenesis, about 20% reduced activity compared to the wild-type enzyme Human gammaherpesvirus 4

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.21 additional information
-
 additional information kinetics, ATP-binding kinetics of wild-type and mutant enzymes at room temperature Human gammaherpesvirus 4
2.7.1.21 0.03
-
 ATP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.041
-
 GTP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.041
-
 ATP recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.043
-
 GTP about, recombinant mutant K297R, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.045
-
 GTP recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.049
-
 CTP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.049
-
 ATP recombinant mutant G294A, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.057
-
 CTP about, recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.069
-
 GTP recombinant mutant G294A, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.072
-
 TTP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.088
-
 CTP recombinant mutant G294A, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.088
-
 ATP recombinant mutant K297R, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.108
-
 CTP recombinant mutant K297R, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 0.127
-
 TTP recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.21 Mg2+ required, most efficient metal ion, can be substituted by Mn2+ or Zn2+ Human gammaherpesvirus 4
2.7.1.21 Mn2+ can partially substitute for Mg2+ Human gammaherpesvirus 4
2.7.1.21 Zn2+ can partially substitute for Mg2+ Human gammaherpesvirus 4

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.21 ATP + thymidine Human gammaherpesvirus 4
-
 ADP + thymidine 5'-phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.21 Human gammaherpesvirus 4
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.21 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) are immobilized on nickel beads and purified Human gammaherpesvirus 4

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.21 ATP + 3'-azido-2',3'-deoxythymidine
-
 Human gammaherpesvirus 4 ADP + 3'-azido-2',3'-deoxythymidine 5'-phosphate
-
 ?
2.7.1.21 ATP + 3'-deoxy-2',3'-didehydro-thymidine
-
 Human gammaherpesvirus 4 ADP + 3'-deoxy-2',3'-didehydro-thymidine 5'-phosphate
-
 ?
2.7.1.21 ATP + aciclovir
-
 Human gammaherpesvirus 4 ADP + aciclovir 5'-phosphate
-
 ?
2.7.1.21 ATP + dideoxyinosine
-
 Human gammaherpesvirus 4 ADP + dideoxyinosine 5'-phosphate
-
 ?
2.7.1.21 ATP + ganciclovir
-
 Human gammaherpesvirus 4 ADP + ganciclovir phosphate
-
 ?
2.7.1.21 ATP + iododeoxyuridine
-
 Human gammaherpesvirus 4 ADP + iododeoxyuridine 5'-phosphate
-
 ?
2.7.1.21 ATP + thymidine
-
 Human gammaherpesvirus 4 ADP + thymidine 5'-phosphate
-
 ?
2.7.1.21 CTP + thymidine
-
 Human gammaherpesvirus 4 CDP + thymidine 5'-phosphate
-
 ?
2.7.1.21 GTP + thymidine
-
 Human gammaherpesvirus 4 GDP + thymidine 5'-phosphate
-
 ?
2.7.1.21 additional information substrate specificity of wild-type and mutant enzymes, overview Human gammaherpesvirus 4 ?
-
 ?
2.7.1.21 TTP + thymidine
-
 Human gammaherpesvirus 4 TDP + thymidine 5'-phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.1.21 TK
-
 Human gammaherpesvirus 4

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.21 37
-
 assay at Human gammaherpesvirus 4

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.21 2 8 ATP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 7
-
 ATP recombinant mutant K297R, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 7
-
 TTP recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 8
-
 GTP recombinant mutant G294A, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 9
-
 CTP recombinant mutant G294A, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 11
-
 ATP recombinant mutant G294A, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 11
-
 TTP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 13.5
-
 CTP about, recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 14
-
 CTP recombinant mutant K297R, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 17
-
 GTP recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 18.5
-
 GTP about, recombinant mutant K297R, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 19
-
 CTP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 21
-
 GTP recombinant wild-type enzyme, pH 7.5, 37°C Human gammaherpesvirus 4
2.7.1.21 24
-
 ATP recombinant mutant T298S, pH 7.5, 37°C Human gammaherpesvirus 4

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.21 7.5
-
 assay at Human gammaherpesvirus 4

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.21 ATP preferred triphosphate cosubstrate, 3 essential residues T298, K297, and G294 in the conserved ATP-binding region, enzyme binding kinetics Human gammaherpesvirus 4
2.7.1.21 CTP less active than ATP Human gammaherpesvirus 4
2.7.1.21 GTP less active tahn ATP Human gammaherpesvirus 4
2.7.1.21 additional information activity with NTP cofactors in descending order ATP, GTP, CTP, TTP Human gammaherpesvirus 4
2.7.1.21 TTP low activity with wild-type and mutant enzymes Human gammaherpesvirus 4