Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.7.1.74 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)

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

Crystallization (Commentary)

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

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 [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

Localization Comment Organism GeneOntology No. Textmining

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Homo sapiens P27707
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms Comment Organism
dCK
-
Homo sapiens

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
ATP
-
Homo sapiens

kcat/KM [mM/s]

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