Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli C41 (DE3) cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
mutant enzyme R104M/D133A in complex with L-thymidine, hanging drop vapor diffusion method, using 0.90-1.5 M trisodium citrate dihydrate and 100 mM HEPES, pH 7.5 | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
R104L/D133A | the double mutant does also accept L- and D-thymidine as substrates | Homo sapiens |
R104M/D133A | the double mutant is a pyrimidine-specific enzyme due to large Km values with purines and does also accept L- and D-thymidine as substrates | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
2'-deoxycytidine | Km below 0.003 mM, wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0057 | - |
2'-deoxycytidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.008 | - |
2'-deoxycytidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0131 | - |
1-beta-D-arabinosylcytosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0161 | - |
gemcitabine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0178 | - |
L-thymidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0243 | - |
D-thymidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0338 | - |
gemcitabine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0506 | - |
1-beta-D-arabinosylcytosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.0562 | - |
gemcitabine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.1146 | - |
2'-deoxyadenosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.1365 | - |
1-beta-D-arabinosylcytosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.138 | - |
L-thymidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.144 | - |
D-thymidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.1623 | - |
2'-deoxyadenosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.231 | - |
2'-deoxyguanosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.04 | - |
2'-deoxyadenosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.865 | - |
2'-deoxyguanosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
2.266 | - |
2'-deoxyguanosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2'-deoxyadenosine | Homo sapiens | - |
ADP + 2'-deoxyadenosine 5'-phosphate | - |
? | |
ATP + 2'-deoxycytidine | Homo sapiens | - |
ADP + 2'-deoxycytidine 5'-phosphate | - |
? | |
ATP + 2'-deoxyguanosine | Homo sapiens | - |
ADP + 2'-deoxyguanosine 5'-phosphate | - |
? | |
ATP + L-thymidine | Homo sapiens | human dCK, in addition to being able to phosphorylate both purines and pyrimidines, has the special ability to accept L-nucleosides as substrates | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P27707 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography, gel filtration | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 1-beta-D-arabinosylcytosine | - |
Homo sapiens | ADP + 1-beta-D-arabinosylcytosine 5'-phosphate | - |
? | |
ATP + 2'-deoxyadenosine | - |
Homo sapiens | ADP + 2'-deoxyadenosine 5'-phosphate | - |
? | |
ATP + 2'-deoxycytidine | - |
Homo sapiens | ADP + 2'-deoxycytidine 5'-phosphate | - |
? | |
ATP + 2'-deoxyguanosine | - |
Homo sapiens | ADP + 2'-deoxyguanosine 5'-phosphate | - |
? | |
ATP + gemcitabine | - |
Homo sapiens | ADP + gemcitabine 5'-phosphate | - |
? | |
ATP + L-thymidine | human dCK, in addition to being able to phosphorylate both purines and pyrimidines, has the special ability to accept L-nucleosides as substrates | Homo sapiens | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
dCK | - |
Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
2'-deoxycytidine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.25 | - |
2'-deoxycytidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.34 | - |
1-beta-D-arabinosylcytosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.39 | - |
gemcitabine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
0.79 | - |
1-beta-D-arabinosylcytosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.33 | - |
L-thymidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.43 | - |
1-beta-D-arabinosylcytosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.73 | - |
2'-deoxyguanosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.74 | - |
D-thymidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.8 | - |
2'-deoxycytidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
2.01 | - |
gemcitabine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
2.13 | - |
2'-deoxyadenosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
2.6 | - |
2'-deoxyguanosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
2.68 | - |
gemcitabine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
3.13 | - |
L-thymidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
3.2 | - |
D-thymidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
3.66 | - |
2'-deoxyguanosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
4.51 | - |
2'-deoxyadenosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
5.72 | - |
2'-deoxyadenosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.9 | - |
2'-deoxyguanosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
1.6 | - |
2'-deoxyguanosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
4.3 | - |
2'-deoxyadenosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
10.5 | - |
1-beta-D-arabinosylcytosine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
11.3 | - |
2'-deoxyguanosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
12.1 | - |
D-thymidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
13.3 | - |
2'-deoxycytidine | kcat/Km above 13.3/sec*mM, wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
15.6 | - |
1-beta-D-arabinosylcytosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
18.6 | - |
2'-deoxyadenosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
22.7 | - |
L-thymidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
24.2 | - |
gemcitabine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
26 | - |
1-beta-D-arabinosylcytosine | wild type enzyme, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
31.3 | - |
2'-deoxycytidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
35.2 | - |
2'-deoxyadenosine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
47.7 | - |
gemcitabine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
59.5 | - |
gemcitabine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
74.7 | - |
L-thymidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
131.7 | - |
D-thymidine | mutant enzyme R104L/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens | |
315.8 | - |
2'-deoxycytidine | mutant enzyme R104M/D133A, in 100 mM Tris, pH 7.5, 100 mM KCl, 10 mM MgCl2 and 1 mM ATP, at 37°C | Homo sapiens |