Application | Comment | Organism |
---|---|---|
medicine | enzyme is a target for antiviral therapy and treatment of EBV-associated malignancies | Human gammaherpesvirus 4 |
Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Human gammaherpesvirus 4 |
Protein Variants | Comment | Organism |
---|---|---|
G294A | site-directed mutagenesis, about 80% reduced activity compared to the wild-type enzyme, no activity with TTP as cofactor | Human gammaherpesvirus 4 |
G294V | site-directed mutagenesis, nearly inactive mutant, no ATP binding | Human gammaherpesvirus 4 |
K297E | site-directed mutagenesis, nearly inactive mutant, no ATP binding | Human gammaherpesvirus 4 |
K297Q | site-directed mutagenesis, nearly inactive mutant, no ATP binding | Human gammaherpesvirus 4 |
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 |
T298A | site-directed mutagenesis, about 90% reduced activity compared to the wild-type enzyme | Human gammaherpesvirus 4 |
T298S | site-directed mutagenesis, about 20% reduced activity compared to the wild-type enzyme | Human gammaherpesvirus 4 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics, ATP-binding kinetics of wild-type and mutant enzymes at room temperature | Human gammaherpesvirus 4 | |
0.03 | - |
ATP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.041 | - |
GTP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.041 | - |
ATP | recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.043 | - |
GTP | about, recombinant mutant K297R, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.045 | - |
GTP | recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.049 | - |
CTP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.049 | - |
ATP | recombinant mutant G294A, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.057 | - |
CTP | about, recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.069 | - |
GTP | recombinant mutant G294A, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.072 | - |
TTP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.088 | - |
CTP | recombinant mutant G294A, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.088 | - |
ATP | recombinant mutant K297R, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.108 | - |
CTP | recombinant mutant K297R, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
0.127 | - |
TTP | recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required, most efficient metal ion, can be substituted by Mn2+ or Zn2+ | Human gammaherpesvirus 4 | |
Mn2+ | can partially substitute for Mg2+ | Human gammaherpesvirus 4 | |
Zn2+ | can partially substitute for Mg2+ | Human gammaherpesvirus 4 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + thymidine | Human gammaherpesvirus 4 | - |
ADP + thymidine 5'-phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Human gammaherpesvirus 4 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 3'-azido-2',3'-deoxythymidine | - |
Human gammaherpesvirus 4 | ADP + 3'-azido-2',3'-deoxythymidine 5'-phosphate | - |
? | |
ATP + 3'-deoxy-2',3'-didehydro-thymidine | - |
Human gammaherpesvirus 4 | ADP + 3'-deoxy-2',3'-didehydro-thymidine 5'-phosphate | - |
? | |
ATP + aciclovir | - |
Human gammaherpesvirus 4 | ADP + aciclovir 5'-phosphate | - |
? | |
ATP + dideoxyinosine | - |
Human gammaherpesvirus 4 | ADP + dideoxyinosine 5'-phosphate | - |
? | |
ATP + ganciclovir | - |
Human gammaherpesvirus 4 | ADP + ganciclovir phosphate | - |
? | |
ATP + iododeoxyuridine | - |
Human gammaherpesvirus 4 | ADP + iododeoxyuridine 5'-phosphate | - |
? | |
ATP + thymidine | - |
Human gammaherpesvirus 4 | ADP + thymidine 5'-phosphate | - |
? | |
CTP + thymidine | - |
Human gammaherpesvirus 4 | CDP + thymidine 5'-phosphate | - |
? | |
GTP + thymidine | - |
Human gammaherpesvirus 4 | GDP + thymidine 5'-phosphate | - |
? | |
additional information | substrate specificity of wild-type and mutant enzymes, overview | Human gammaherpesvirus 4 | ? | - |
? | |
TTP + thymidine | - |
Human gammaherpesvirus 4 | TDP + thymidine 5'-phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
TK | - |
Human gammaherpesvirus 4 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Human gammaherpesvirus 4 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | 8 | ATP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
7 | - |
ATP | recombinant mutant K297R, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
7 | - |
TTP | recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
8 | - |
GTP | recombinant mutant G294A, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
9 | - |
CTP | recombinant mutant G294A, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
11 | - |
ATP | recombinant mutant G294A, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
11 | - |
TTP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
13.5 | - |
CTP | about, recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
14 | - |
CTP | recombinant mutant K297R, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
17 | - |
GTP | recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
18.5 | - |
GTP | about, recombinant mutant K297R, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
19 | - |
CTP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
21 | - |
GTP | recombinant wild-type enzyme, pH 7.5, 37°C | Human gammaherpesvirus 4 | |
24 | - |
ATP | recombinant mutant T298S, pH 7.5, 37°C | Human gammaherpesvirus 4 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Human gammaherpesvirus 4 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | preferred triphosphate cosubstrate, 3 essential residues T298, K297, and G294 in the conserved ATP-binding region, enzyme binding kinetics | Human gammaherpesvirus 4 | |
CTP | less active than ATP | Human gammaherpesvirus 4 | |
GTP | less active tahn ATP | Human gammaherpesvirus 4 | |
additional information | activity with NTP cofactors in descending order ATP, GTP, CTP, TTP | Human gammaherpesvirus 4 | |
TTP | low activity with wild-type and mutant enzymes | Human gammaherpesvirus 4 |