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Literature summary extracted from

  • Hazra, S.; Sabini, E.; Ort, S.; Konrad, M.; Lavie, A.
    Extending thymidine kinase activity to the catalytic repertoire of human deoxycytidine kinase (2009), Biochemistry, 48, 1256-1263.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.74 expressed in Escherichia coli C41 (DE3) cells Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.1.74 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

EC Number Protein Variants Comment Organism
2.7.1.74 R104L/D133A the double mutant does also accept L- and D-thymidine as substrates Homo sapiens
2.7.1.74 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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

EC Number Localization Comment Organism GeneOntology No. Textmining

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.74 ATP + 2'-deoxyadenosine Homo sapiens
-
ADP + 2'-deoxyadenosine 5'-phosphate
-
?
2.7.1.74 ATP + 2'-deoxycytidine Homo sapiens
-
ADP + 2'-deoxycytidine 5'-phosphate
-
?
2.7.1.74 ATP + 2'-deoxyguanosine Homo sapiens
-
ADP + 2'-deoxyguanosine 5'-phosphate
-
?
2.7.1.74 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

EC Number Organism UniProt Comment Textmining
2.7.1.74 Homo sapiens P27707
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.74 Ni-NTA column chromatography, gel filtration Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.74 ATP + 1-beta-D-arabinosylcytosine
-
Homo sapiens ADP + 1-beta-D-arabinosylcytosine 5'-phosphate
-
?
2.7.1.74 ATP + 2'-deoxyadenosine
-
Homo sapiens ADP + 2'-deoxyadenosine 5'-phosphate
-
?
2.7.1.74 ATP + 2'-deoxycytidine
-
Homo sapiens ADP + 2'-deoxycytidine 5'-phosphate
-
?
2.7.1.74 ATP + 2'-deoxyguanosine
-
Homo sapiens ADP + 2'-deoxyguanosine 5'-phosphate
-
?
2.7.1.74 ATP + gemcitabine
-
Homo sapiens ADP + gemcitabine 5'-phosphate
-
?
2.7.1.74 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

EC Number Synonyms Comment Organism
2.7.1.74 dCK
-
Homo sapiens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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

EC Number Cofactor Comment Organism Structure
2.7.1.74 ATP
-
Homo sapiens

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 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
2.7.1.74 131.7
-
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
2.7.1.74 315.8
-
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