BRENDA - Enzyme Database show

Deoxycytidine kinase. II. Purification and general properties of the calf thymus enzyme

Durham, J.P.; Ives, D.H.; J. Biol. Chem. 245, 2276-2284 (1970)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.74
bovine serum albumin
activation
Bos taurus
2.7.1.74
dithiothreitol
activation
Bos taurus
General Stability
EC Number
General Stability
Organism
2.7.1.74
2-mercaptoethanol stabilizes
Bos taurus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.74
ADP
not
Bos taurus
2.7.1.74
CMP
not
Bos taurus
2.7.1.74
cytidine
cytosine arabinoside as substrate
Bos taurus
2.7.1.74
cytosine arabinoside
deoxyguanosine as substrate
Bos taurus
2.7.1.74
dAMP
not
Bos taurus
2.7.1.74
dCDP
-
Bos taurus
2.7.1.74
dCMP
-
Bos taurus
2.7.1.74
dCTP
reversible by dTTP; reversible by UTP, dUTP
Bos taurus
2.7.1.74
deoxyadenosine
cytosine arabinoside or deoxyguanosine as substrate
Bos taurus
2.7.1.74
deoxycytidine
cytosine arabinoside as substrate; deoxyadenosine as substrate; deoxyguanosine as substrate
Bos taurus
2.7.1.74
deoxyguanosine
cytosine arabinoside as substrate
Bos taurus
2.7.1.74
deoxythymidine
cytosolic isozyme I, not
Bos taurus
2.7.1.74
dTDP
-
Bos taurus
2.7.1.74
dUMP
not
Bos taurus
2.7.1.74
GDP
not
Bos taurus
2.7.1.74
GSSG
thiol reagents protect or reverse
Bos taurus
2.7.1.74
HgCl2
strong, thiol reagents protect
Bos taurus
2.7.1.74
additional information
little or no inhibition by AMP, dTMP, adenosine, guanosine; little or no inhibition by deoxyuridine
Bos taurus
2.7.1.74
N-ethylmaleimide
thiol reagents protect or reverse
Bos taurus
2.7.1.74
p-chloromercuribenzoate
thiol reagents protect or reverse
Bos taurus
2.7.1.74
UDP
-
Bos taurus
2.7.1.74
UMP
not
Bos taurus
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.74
Ca2+
about 55% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
CdCl2
activation, in the absence of dithiothreitol
Bos taurus
2.7.1.74
Co2+
about 40% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Fe2+
about 45% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Mg2+
-
Bos taurus
2.7.1.74
Mn2+
about 80% as efficient as Mg2+; requirement
Bos taurus
2.7.1.74
additional information
no activation by Ba2+, Cu2+
Bos taurus
2.7.1.74
Ni2+
about 20% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Sr2+
about 15% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Zn2+
about 40% as efficient as Mg2+; activation
Bos taurus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.74
56000
-
gel filtration, equilibrium density and sucrose density gradient centrifugation
Bos taurus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.1.74
Bos taurus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.1.74
-
Bos taurus
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
2.7.1.74
thymus
-
Bos taurus
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.74
0.012
-
37°C, pH 8.0
Bos taurus
Storage Stability
EC Number
Storage Stability
Organism
2.7.1.74
-30°C, several months in the presence of 2-mercaptoethanol
Bos taurus
2.7.1.74
4°C, if purified in the absence of 2-mercaptoethanol it loses most of its activity within 2 days, DTT restores
Bos taurus
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 + 2'-deoxycytidine
in the presence of DTT less effective than dUTP, UTP, dTTP or GTP, without DTT only less effective than dUTP
641931
Bos taurus
ADP + 5'-dCMP
-
641931
Bos taurus
?
2.7.1.74
dTTP + 2'-deoxycytidine
-
641931
Bos taurus
dTDP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
dUTP + 2'-deoxycytidine
best phosphate donor
641931
Bos taurus
dUDP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
GTP + 2'-deoxycytidine
-
641931
Bos taurus
GDP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
NTP + cytosine arabinoside
-
641931
Bos taurus
NDP + cytosine arabinoside 5'-monophosphate
-
-
-
?
2.7.1.74
NTP + deoxyadenosine
-
641931
Bos taurus
NDP + dAMP
-
-
-
?
2.7.1.74
NTP + deoxycytidine
-
641931
Bos taurus
NDP + dCMP
-
641931
Bos taurus
?
2.7.1.74
NTP + deoxyguanosine
-
641931
Bos taurus
NDP + dGMP
-
-
-
?
2.7.1.74
UTP + 2'-deoxycytidine
-
641931
Bos taurus
UDP + 2'-deoxy-CMP
-
-
-
?
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.74
37
-
assay at
Bos taurus
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.1.74
22
-
at least 24 h, in the presence of dithiothreitol
Bos taurus
2.7.1.74
60
-
dCTP enhances heat stability at pH 6, dithiothreitol slightly, not deoxycytidine or other nucleosides; t1/2: 7.9 min at pH 8, t1/2: 2 min at pH 6, unstable at pH 10
Bos taurus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.74
6
10
broad plateau, with very slight optima at pH 6 and 10
Bos taurus
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
2.7.1.74
6
-
t1/2: 2 min at 60°C
Bos taurus
2.7.1.74
8
-
moderately stable, t1/2: 7.9 min at 60°C
Bos taurus
2.7.1.74
10
-
unstable at 60°C
Bos taurus
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
2.7.1.74
bovine serum albumin
activation
Bos taurus
2.7.1.74
dithiothreitol
activation
Bos taurus
General Stability (protein specific)
EC Number
General Stability
Organism
2.7.1.74
2-mercaptoethanol stabilizes
Bos taurus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.7.1.74
ADP
not
Bos taurus
2.7.1.74
CMP
not
Bos taurus
2.7.1.74
cytidine
cytosine arabinoside as substrate
Bos taurus
2.7.1.74
cytosine arabinoside
deoxyguanosine as substrate
Bos taurus
2.7.1.74
dAMP
not
Bos taurus
2.7.1.74
dCDP
-
Bos taurus
2.7.1.74
dCMP
-
Bos taurus
2.7.1.74
dCTP
reversible by dTTP; reversible by UTP, dUTP
Bos taurus
2.7.1.74
deoxyadenosine
cytosine arabinoside or deoxyguanosine as substrate
Bos taurus
2.7.1.74
deoxycytidine
cytosine arabinoside as substrate; deoxyadenosine as substrate; deoxyguanosine as substrate
Bos taurus
2.7.1.74
deoxyguanosine
cytosine arabinoside as substrate
Bos taurus
2.7.1.74
deoxythymidine
cytosolic isozyme I, not
Bos taurus
2.7.1.74
dTDP
-
Bos taurus
2.7.1.74
dUMP
not
Bos taurus
2.7.1.74
GDP
not
Bos taurus
2.7.1.74
GSSG
thiol reagents protect or reverse
Bos taurus
2.7.1.74
HgCl2
strong, thiol reagents protect
Bos taurus
2.7.1.74
additional information
little or no inhibition by AMP, dTMP, adenosine, guanosine; little or no inhibition by deoxyuridine
Bos taurus
2.7.1.74
N-ethylmaleimide
thiol reagents protect or reverse
Bos taurus
2.7.1.74
p-chloromercuribenzoate
thiol reagents protect or reverse
Bos taurus
2.7.1.74
UDP
-
Bos taurus
2.7.1.74
UMP
not
Bos taurus
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.1.74
Ca2+
about 55% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
CdCl2
activation, in the absence of dithiothreitol
Bos taurus
2.7.1.74
Co2+
about 40% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Fe2+
about 45% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Mg2+
-
Bos taurus
2.7.1.74
Mn2+
about 80% as efficient as Mg2+; requirement
Bos taurus
2.7.1.74
additional information
no activation by Ba2+, Cu2+
Bos taurus
2.7.1.74
Ni2+
about 20% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Sr2+
about 15% as efficient as Mg2+; activation
Bos taurus
2.7.1.74
Zn2+
about 40% as efficient as Mg2+; activation
Bos taurus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.1.74
56000
-
gel filtration, equilibrium density and sucrose density gradient centrifugation
Bos taurus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.1.74
-
Bos taurus
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
2.7.1.74
thymus
-
Bos taurus
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.1.74
0.012
-
37°C, pH 8.0
Bos taurus
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
2.7.1.74
-30°C, several months in the presence of 2-mercaptoethanol
Bos taurus
2.7.1.74
4°C, if purified in the absence of 2-mercaptoethanol it loses most of its activity within 2 days, DTT restores
Bos taurus
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 + 2'-deoxycytidine
in the presence of DTT less effective than dUTP, UTP, dTTP or GTP, without DTT only less effective than dUTP
641931
Bos taurus
ADP + 5'-dCMP
-
641931
Bos taurus
?
2.7.1.74
dTTP + 2'-deoxycytidine
-
641931
Bos taurus
dTDP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
dUTP + 2'-deoxycytidine
best phosphate donor
641931
Bos taurus
dUDP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
GTP + 2'-deoxycytidine
-
641931
Bos taurus
GDP + 2'-deoxy-CMP
-
-
-
?
2.7.1.74
NTP + cytosine arabinoside
-
641931
Bos taurus
NDP + cytosine arabinoside 5'-monophosphate
-
-
-
?
2.7.1.74
NTP + deoxyadenosine
-
641931
Bos taurus
NDP + dAMP
-
-
-
?
2.7.1.74
NTP + deoxycytidine
-
641931
Bos taurus
NDP + dCMP
-
641931
Bos taurus
?
2.7.1.74
NTP + deoxyguanosine
-
641931
Bos taurus
NDP + dGMP
-
-
-
?
2.7.1.74
UTP + 2'-deoxycytidine
-
641931
Bos taurus
UDP + 2'-deoxy-CMP
-
-
-
?
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.7.1.74
37
-
assay at
Bos taurus
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.1.74
22
-
at least 24 h, in the presence of dithiothreitol
Bos taurus
2.7.1.74
60
-
dCTP enhances heat stability at pH 6, dithiothreitol slightly, not deoxycytidine or other nucleosides; t1/2: 7.9 min at pH 8, t1/2: 2 min at pH 6, unstable at pH 10
Bos taurus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.7.1.74
6
10
broad plateau, with very slight optima at pH 6 and 10
Bos taurus
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
2.7.1.74
6
-
t1/2: 2 min at 60°C
Bos taurus
2.7.1.74
8
-
moderately stable, t1/2: 7.9 min at 60°C
Bos taurus
2.7.1.74
10
-
unstable at 60°C
Bos taurus