BRENDA - Enzyme Database show

Extending thymidine kinase activity to the catalytic repertoire of human deoxycytidine kinase

Hazra, S.; Sabini, E.; Ort, S.; Konrad, M.; Lavie, A.; Biochemistry 48, 1256-1263 (2009)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.74
expressed in Escherichia coli C41 (DE3) cells
Homo sapiens
Crystallization (Commentary)
EC Number
Crystallization
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
Engineering
EC Number
Amino acid exchange
Commentary
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
Commentary
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
Commentary
Organism
GeneOntology No.
Textmining
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.74
Homo sapiens
P27707
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.74
Ni-NTA column chromatography, gel filtration
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.74
ATP + 1-beta-D-arabinosylcytosine
-
702271
Homo sapiens
ADP + 1-beta-D-arabinosylcytosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxyadenosine
-
702271
Homo sapiens
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxycytidine
-
702271
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxyguanosine
-
702271
Homo sapiens
ADP + 2'-deoxyguanosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + gemcitabine
-
702271
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
702271
Homo sapiens
?
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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
Commentary
Organism
Structure
2.7.1.74
ATP
-
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
expressed in Escherichia coli C41 (DE3) cells
Homo sapiens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.74
ATP
-
Homo sapiens
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
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
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
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] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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 (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
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
?
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
Ni-NTA column chromatography, gel filtration
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.1.74
ATP + 1-beta-D-arabinosylcytosine
-
702271
Homo sapiens
ADP + 1-beta-D-arabinosylcytosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxyadenosine
-
702271
Homo sapiens
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxycytidine
-
702271
Homo sapiens
ADP + 2'-deoxycytidine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxyguanosine
-
702271
Homo sapiens
ADP + 2'-deoxyguanosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + gemcitabine
-
702271
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
702271
Homo sapiens
?
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
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
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
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