BRENDA - Enzyme Database show

Mimicking phosphorylation of Ser-74 on human deoxycytidine kinase selectively increases catalytic activity for dC and dC analogues

McSorley, T.; Ort, S.; Hazra, S.; Lavie, A.; Konrad, M.; FEBS Lett. 582, 720-724 (2008)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.74
expression in Escherichia coli and in HEK-293 cell
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.74
S74A
6-8fold lower activity than wild-type
Homo sapiens
2.7.1.74
S74D
2-4fold lower kcat values
Homo sapiens
2.7.1.74
S74E
mutation mimicking phosphorylation, results in 11fold increase in kcat value for deoxycytidine. Mutation has no effect on cellular localization
Homo sapiens
2.7.1.74
S74Q
no significant change in kcat for any of the substrates tested
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.74
0.0017
-
2'-deoxycytidine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.0059
-
1-beta-D-arabinofuranosylcytosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.0062
-
2',2'-difluorodeoxycytidine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.0089
-
2'-deoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.015
-
2',2'-difluorodeoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.024
-
1-beta-D-arabinofuranosylcytosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.1169
-
2'-deoxyadenosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.2458
-
2'-deoxyguanosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.518
-
2'-deoxyadenosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.602
-
2'-deoxyguanosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.1.74
nucleus
main localization of both wild-type and mutant S74E mimicking phosphorylated enzyme
Homo sapiens
5634
-
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.74
30000
-
x * 30000, SDS-PAGE
Homo sapiens
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.74
Homo sapiens
P27707
-
-
Posttranslational Modification
EC Number
Posttranslational Modification
Commentary
Organism
2.7.1.74
phosphoprotein
phosphorylation at residue S74
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-arabinofuranosylcytosine
-
692364
Homo sapiens
ADP + 1-beta-D-arabinofuranosylcytosine 5'-phosphate
-
-
-
-
2.7.1.74
ATP + 2',2'-difluorodeoxycytidine
-
692364
Homo sapiens
ADP + 2',2'-difluorodeoxycytidine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxyadenosine
-
692364
Homo sapiens
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxycytidine
-
692364
Homo sapiens
ADP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
ATP + 2'-deoxyguanosine
-
692364
Homo sapiens
ADP + 2'-deoxyguanosine 5'-phosphate
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
2.7.1.74
?
x * 30000, SDS-PAGE
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
mutant S74A, pH 7.5, 25°C; mutant S74Q, pH 7.5, 25°C; wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.16
-
2'-deoxycytidine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.3
-
1-beta-D-arabinofuranosylcytosine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.32
-
1-beta-D-arabinofuranosylcytosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.33
-
1-beta-D-arabinofuranosylcytosine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.36
-
2'-deoxyguanosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.44
-
2',2'-difluorodeoxycytidine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.45
-
2',2'-difluorodeoxycytidine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.45
-
2'-deoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.5
-
2',2'-difluorodeoxycytidine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.57
-
1-beta-D-arabinofuranosylcytosine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.57
-
2'-deoxyguanosine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.65
-
2'-deoxyguanosine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.68
-
2'-deoxyguanosine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.69
-
2'-deoxyadenosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.73
-
2'-deoxyguanosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.77
-
2'-deoxyadenosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.9
-
2',2'-difluorodeoxycytidine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.9
-
2'-deoxyadenosine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1
-
2'-deoxyadenosine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1.06
-
1-beta-D-arabinofuranosylcytosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1.12
-
2'-deoxyadenosine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1.21
-
2',2'-difluorodeoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
expression in Escherichia coli and in HEK-293 cell
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.74
S74A
6-8fold lower activity than wild-type
Homo sapiens
2.7.1.74
S74D
2-4fold lower kcat values
Homo sapiens
2.7.1.74
S74E
mutation mimicking phosphorylation, results in 11fold increase in kcat value for deoxycytidine. Mutation has no effect on cellular localization
Homo sapiens
2.7.1.74
S74Q
no significant change in kcat for any of the substrates tested
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.74
0.0017
-
2'-deoxycytidine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.0059
-
1-beta-D-arabinofuranosylcytosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.0062
-
2',2'-difluorodeoxycytidine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.0089
-
2'-deoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.015
-
2',2'-difluorodeoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.024
-
1-beta-D-arabinofuranosylcytosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.1169
-
2'-deoxyadenosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.2458
-
2'-deoxyguanosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.518
-
2'-deoxyadenosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.602
-
2'-deoxyguanosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.1.74
nucleus
main localization of both wild-type and mutant S74E mimicking phosphorylated enzyme
Homo sapiens
5634
-
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.74
30000
-
x * 30000, SDS-PAGE
Homo sapiens
Posttranslational Modification (protein specific)
EC Number
Posttranslational Modification
Commentary
Organism
2.7.1.74
phosphoprotein
phosphorylation at residue S74
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-arabinofuranosylcytosine
-
692364
Homo sapiens
ADP + 1-beta-D-arabinofuranosylcytosine 5'-phosphate
-
-
-
-
2.7.1.74
ATP + 2',2'-difluorodeoxycytidine
-
692364
Homo sapiens
ADP + 2',2'-difluorodeoxycytidine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxyadenosine
-
692364
Homo sapiens
ADP + 2'-deoxyadenosine 5'-phosphate
-
-
-
?
2.7.1.74
ATP + 2'-deoxycytidine
-
692364
Homo sapiens
ADP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
ATP + 2'-deoxyguanosine
-
692364
Homo sapiens
ADP + 2'-deoxyguanosine 5'-phosphate
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.1.74
?
x * 30000, SDS-PAGE
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
mutant S74A, pH 7.5, 25°C; mutant S74Q, pH 7.5, 25°C; wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.16
-
2'-deoxycytidine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.3
-
1-beta-D-arabinofuranosylcytosine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.32
-
1-beta-D-arabinofuranosylcytosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.33
-
1-beta-D-arabinofuranosylcytosine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.36
-
2'-deoxyguanosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.44
-
2',2'-difluorodeoxycytidine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.45
-
2',2'-difluorodeoxycytidine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.45
-
2'-deoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.5
-
2',2'-difluorodeoxycytidine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.57
-
1-beta-D-arabinofuranosylcytosine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.57
-
2'-deoxyguanosine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.65
-
2'-deoxyguanosine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.68
-
2'-deoxyguanosine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.69
-
2'-deoxyadenosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.73
-
2'-deoxyguanosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.77
-
2'-deoxyadenosine
wild-type, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.9
-
2',2'-difluorodeoxycytidine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
0.9
-
2'-deoxyadenosine
mutant S74Q, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1
-
2'-deoxyadenosine
mutant S74D, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1.06
-
1-beta-D-arabinofuranosylcytosine
mutant S74E, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1.12
-
2'-deoxyadenosine
mutant S74A, pH 7.5, 25°C
Homo sapiens
2.7.1.74
1.21
-
2',2'-difluorodeoxycytidine
mutant S74E, pH 7.5, 25°C
Homo sapiens