BRENDA - Enzyme Database show

Post-translational phosphorylation of serine 74 of human deoxycytidine kinase favors the enzyme adopting the open conformation making it competent for nucleoside binding and release

Hazra, S.; Szewczak, A.; Ort, S.; Konrad, M.; Lavie, A.; Biochemistry 50, 2870-2880 (2011)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.1.74
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain C41(DE3)
Homo sapiens
Crystallization (Commentary)
EC Number
Crystallization
Organism
2.7.1.74
enzyme mutant C9S/C45S/C59S/S74E/R104M/D133A/C146S in complex with L-dT and UDP, hanging drop vapour diffusion method, mixing of 0.001 ml of 9-15 mg/ml protein mixed with 0.001 ml reservoir solution containing 0.9-1.5 M trisodium citrate dihydrate and 100 mM Tris, pH 7.5, room temperature, X-ray diffraction structure determination and analysis at 1.9-2.5 A resolution
Homo sapiens
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.74
C9S/C45S/C59S/S74E/R104M/D133A/C146S
site-directed mutagenesis, crystallization mutant
Homo sapiens
2.7.1.74
D133A
site-directed mutagenesis, the mutation renders the enzyme capable of thymidine binding
Homo sapiens
2.7.1.74
R104M
site-directed mutagenesis, the mutation renders the enzyme capable of thymidine binding
Homo sapiens
2.7.1.74
S74E
site-directed mutagenesis, replacing the serine with a glutamic acid mimic phosphorylation of Ser74, the enzyme containing the S74E mutation adopts the open state, but wild-type dCK can adopt the open state also in the absence of the S74E mutation
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.74
additional information
-
additional information
kinetic properties of the enzyme are modulated in vivo by phosphorylation of Ser74, mechanism, overview
Homo sapiens
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.74
Mg2+
required
Homo sapiens
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 + deoxycytidine
Homo sapiens
-
ADP + dCMP
-
-
?
2.7.1.74
UTP + deoxycytidine
Homo sapiens
-
UDP + dCMP
-
-
?
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
kinetic properties of the enzyme are modulated in vivo by phosphorylation of Ser74, conformational changes and mechanism, overview
Homo sapiens
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.74
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain C41(DE3) by nickel affinity chromatography and 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 + deoxycytidine
-
721636
Homo sapiens
ADP + dCMP
-
-
-
?
2.7.1.74
UTP + deoxycytidine
-
721636
Homo sapiens
UDP + dCMP
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.74
37
-
assay at
Homo sapiens
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.74
7.5
-
assay at
Homo sapiens
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.74
ATP
-
Homo sapiens
2.7.1.74
UTP
-
Homo sapiens
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain C41(DE3)
Homo sapiens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.1.74
ATP
-
Homo sapiens
2.7.1.74
UTP
-
Homo sapiens
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
2.7.1.74
enzyme mutant C9S/C45S/C59S/S74E/R104M/D133A/C146S in complex with L-dT and UDP, hanging drop vapour diffusion method, mixing of 0.001 ml of 9-15 mg/ml protein mixed with 0.001 ml reservoir solution containing 0.9-1.5 M trisodium citrate dihydrate and 100 mM Tris, pH 7.5, room temperature, X-ray diffraction structure determination and analysis at 1.9-2.5 A resolution
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.7.1.74
C9S/C45S/C59S/S74E/R104M/D133A/C146S
site-directed mutagenesis, crystallization mutant
Homo sapiens
2.7.1.74
D133A
site-directed mutagenesis, the mutation renders the enzyme capable of thymidine binding
Homo sapiens
2.7.1.74
R104M
site-directed mutagenesis, the mutation renders the enzyme capable of thymidine binding
Homo sapiens
2.7.1.74
S74E
site-directed mutagenesis, replacing the serine with a glutamic acid mimic phosphorylation of Ser74, the enzyme containing the S74E mutation adopts the open state, but wild-type dCK can adopt the open state also in the absence of the S74E mutation
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.1.74
additional information
-
additional information
kinetic properties of the enzyme are modulated in vivo by phosphorylation of Ser74, mechanism, overview
Homo sapiens
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.74
Mg2+
required
Homo sapiens
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 + deoxycytidine
Homo sapiens
-
ADP + dCMP
-
-
?
2.7.1.74
UTP + deoxycytidine
Homo sapiens
-
UDP + dCMP
-
-
?
Posttranslational Modification (protein specific)
EC Number
Posttranslational Modification
Commentary
Organism
2.7.1.74
phosphoprotein
kinetic properties of the enzyme are modulated in vivo by phosphorylation of Ser74, conformational changes and mechanism, overview
Homo sapiens
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain C41(DE3) by nickel affinity chromatography and 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 + deoxycytidine
-
721636
Homo sapiens
ADP + dCMP
-
-
-
?
2.7.1.74
UTP + deoxycytidine
-
721636
Homo sapiens
UDP + dCMP
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.74
37
-
assay at
Homo sapiens
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.74
7.5
-
assay at
Homo sapiens
General Information
EC Number
General Information
Commentary
Organism
2.7.1.74
additional information
structure-function analysis, comparison of wild-type enzyme and mutant S74E
Homo sapiens
2.7.1.74
physiological function
phosphorylation of Ser74 might be a mechanism for regulating the specificity of the kinase to favor deoxycytidine
Homo sapiens
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.7.1.74
additional information
structure-function analysis, comparison of wild-type enzyme and mutant S74E
Homo sapiens
2.7.1.74
physiological function
phosphorylation of Ser74 might be a mechanism for regulating the specificity of the kinase to favor deoxycytidine
Homo sapiens