BRENDA - Enzyme Database
show all sequences of 2.7.4.25

Structural and functional consequences of single amino acid substitutions in the pyrimidine base binding pocket of Escherichia coli CMP kinase

Ofiteru, A.; Bucurenci, N.; Alexov, E.; Bertrand, T.; Briozzo, P.; Munier-Lehmann, H.; Gilles, A.M.; FEBS J. 274, 3363-3373 (2007)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli BL21(DE3) cells
Escherichia coli K-12
expression of wild-type and mutant enzymes
Escherichia coli
Crystallization (Commentary)
Crystallization
Organism
mutant enzyme R188M is crystallized either alone or in complex with dCMP, hanging drop vapor diffusion method, using ammonium sulfate as a precipitant (1.3 M in the case of enzyme alone, and 1.7 M for the R188M-dCMP complex)
Escherichia coli K-12
purified recombinant mutant R188M free or in complex with CMP, X-ray diffraction structure determination and analysis at 1.9 A and 2.8 A resolution, respectively
Escherichia coli
Engineering
Amino acid exchange
Commentary
Organism
D132A
the mutant shows reduced activity compared to the wild type enzyme, the mutation has the most dramatic consequences on the protein stability as Tm decreases by 9°C in comparison with the wild type protein, an increase in the specificity of the D132A variant for UMP over CMP and dCMP is observed
Escherichia coli K-12
D132A
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
D132H
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
D132H
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP, the D132H variant does not introduce charge reversal, because His is calculated to be deprotonated
Escherichia coli
D132N
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
D132N
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli K-12
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
R110M
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
R110M
the side chains of Arg110 cannot establish hydrogen bonds with UMP, and its substitution by hydrophobic amino acids simultaneously affects the Km of CMP/dCMP and the kcat value
Escherichia coli
R188M
the mutation does not affect enzyme stability but dramatically decreases the kcat_Km ratio compared with wild type enzyme, the mutant has no activity towards UMP
Escherichia coli K-12
R188M
replacement of Arg188 with Met does not affect enzyme stability but dramatically decreases the kcat/Km ratio compared to the wild-type enzyme
Escherichia coli
S36A
the S36A substitution mainly changes the Km value for the two natural substrates, which increases by a factor of 70 (CMP) and 37 (dCMP) compared with the parent molecule and decreases kcat of 1.6fold (CMP) and 7.4fold (dCMP) with respect to the wild type enzyme
Escherichia coli K-12
S36A
the side chains of Arg110 cannot establish hydrogen bonds with UMP, and its substitution by hydrophobic amino acids simultaneously affects the Km of CMP/dCMP and the kcat value
Escherichia coli
General Stability
General Stability
Organism
four key amino acids interacting with the pyrimidine ring of CMP, Ser36, Asp132, Arg110 and Arg188, contribute to the stability, catalysis and substrate specificity of Escherichia coli CMP kinase
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics and substrate specificities of wild-type and mutant enzymes, overview
Escherichia coli
0.035
-
CMP
wild-type enzyme
Escherichia coli
0.035
-
CMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.038
-
CMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0394
-
dCMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.055
-
dCMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.08
-
dCMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.09
-
dCMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.094
-
dCMP
wild-type enzyme
Escherichia coli
0.77
-
dCMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.93
-
UMP
wild-type enzyme
Escherichia coli
0.93
-
UMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1
-
CMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.3
-
CMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.8
-
dCMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.9
-
UMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2.5
-
CMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2.6
-
CMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2.9
-
CMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
3.5
-
dCMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
3.9
-
UMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
5.4
-
UMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
7.3
-
dCMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
7.9
-
UMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
8
-
UMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
11.3
-
UMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
20.2
-
CMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
required
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
24700
-
calculated from amino acid sequence
Escherichia coli K-12
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + CMP
Escherichia coli
-
ADP + CDP
-
-
r
ATP + dCMP
Escherichia coli
-
ADP + dCDP
-
-
r
additional information
Escherichia coli
bacterial CMP kinases are specific for CMP and dCMP, whereas the related eukaryotic NMP kinase phosphorylates CMP and UMP with similar efficiency. Bacterial CMP kinase has a narrower NMP-binding pocket and a hydrogen-bonding network involving the pyrimidine moiety specific for the cytosine nucleobase compared to eukaryotic UMP?CMP kinases
?
-
-
-
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
P0A6I0
-
-
Escherichia coli K-12
P0A6I0
strain CJ236
-
Purification (Commentary)
Commentary
Organism
-
Escherichia coli K-12
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
-
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + CMP
-
692282
Escherichia coli
ADP + CDP
-
-
-
r
ATP + CMP
the enzyme is specific for CMP
692282
Escherichia coli K-12
ADP + CDP
-
-
-
?
ATP + dCMP
-
692282
Escherichia coli
ADP + dCDP
-
-
-
r
ATP + dCMP
the enzyme is specific for dCMP
692282
Escherichia coli K-12
ADP + dCDP
-
-
-
?
ATP + UMP
UMP is a poor substrate
692282
Escherichia coli
ADP + UDP
-
-
-
r
ATP + UMP
UMP is a weak substrate
692282
Escherichia coli K-12
ADP + UDP
-
-
-
?
additional information
bacterial CMP kinases are specific for CMP and dCMP, whereas the related eukaryotic NMP kinase phosphorylates CMP and UMP with similar efficiency. Bacterial CMP kinase has a narrower NMP-binding pocket and a hydrogen-bonding network involving the pyrimidine moiety specific for the cytosine nucleobase compared to eukaryotic UMP?CMP kinases
692282
Escherichia coli
?
-
-
-
-
additional information
four key amino acids interacting with the pyrimidine ring of CMP, Ser36, Asp132, Arg110 and Arg188, contribute to the stability, catalysis and substrate specificity of Escherichia coli CMP kinase. D132 is involved in a complicated network of interactions, the strongest being with R110, and also R110 is involved in many interactions, but mainly with D132. Substrate specificities of wild-type and mutant enzymes, overview
692282
Escherichia coli
?
-
-
-
-
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
additional information
-
thermal stability of CMP kinase genetic variants, overview
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.013
-
UMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.04
-
dCMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.05
-
dCMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.054
-
UMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.06
-
dCMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.069
-
CMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.12
-
CMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.15
-
dCMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.23
-
CMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.45
-
UMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.57
-
UMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.73
-
dCMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.82
-
UMP
wild-type enzyme
Escherichia coli
0.82
-
UMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.4
-
CMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
4.1
-
CMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
8.3
-
UMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
9.9
-
UMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
14.5
-
dCMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
21.1
-
dCMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
22.4
-
CMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
63
-
CMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
103
-
CMP
wild-type enzyme
Escherichia coli
103
-
CMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
108
-
dCMP
wild-type enzyme
Escherichia coli
108
-
dCMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
Cofactor
Cofactor
Commentary
Organism
Structure
ATP
-
Escherichia coli K-12
ATP
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21(DE3) cells
Escherichia coli K-12
expression of wild-type and mutant enzymes
Escherichia coli
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
ATP
-
Escherichia coli
ATP
-
Escherichia coli K-12
Crystallization (Commentary) (protein specific)
Crystallization
Organism
mutant enzyme R188M is crystallized either alone or in complex with dCMP, hanging drop vapor diffusion method, using ammonium sulfate as a precipitant (1.3 M in the case of enzyme alone, and 1.7 M for the R188M-dCMP complex)
Escherichia coli K-12
purified recombinant mutant R188M free or in complex with CMP, X-ray diffraction structure determination and analysis at 1.9 A and 2.8 A resolution, respectively
Escherichia coli
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D132A
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
D132A
the mutant shows reduced activity compared to the wild type enzyme, the mutation has the most dramatic consequences on the protein stability as Tm decreases by 9°C in comparison with the wild type protein, an increase in the specificity of the D132A variant for UMP over CMP and dCMP is observed
Escherichia coli K-12
D132H
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP, the D132H variant does not introduce charge reversal, because His is calculated to be deprotonated
Escherichia coli
D132H
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
D132N
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
D132N
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli K-12
R110M
the side chains of Arg110 cannot establish hydrogen bonds with UMP, and its substitution by hydrophobic amino acids simultaneously affects the Km of CMP/dCMP and the kcat value
Escherichia coli
R110M
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
R188M
replacement of Arg188 with Met does not affect enzyme stability but dramatically decreases the kcat/Km ratio compared to the wild-type enzyme
Escherichia coli
R188M
the mutation does not affect enzyme stability but dramatically decreases the kcat_Km ratio compared with wild type enzyme, the mutant has no activity towards UMP
Escherichia coli K-12
S36A
the side chains of Arg110 cannot establish hydrogen bonds with UMP, and its substitution by hydrophobic amino acids simultaneously affects the Km of CMP/dCMP and the kcat value
Escherichia coli
S36A
the S36A substitution mainly changes the Km value for the two natural substrates, which increases by a factor of 70 (CMP) and 37 (dCMP) compared with the parent molecule and decreases kcat of 1.6fold (CMP) and 7.4fold (dCMP) with respect to the wild type enzyme
Escherichia coli K-12
General Stability (protein specific)
General Stability
Organism
four key amino acids interacting with the pyrimidine ring of CMP, Ser36, Asp132, Arg110 and Arg188, contribute to the stability, catalysis and substrate specificity of Escherichia coli CMP kinase
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
kinetics and substrate specificities of wild-type and mutant enzymes, overview
Escherichia coli
0.035
-
CMP
wild-type enzyme
Escherichia coli
0.035
-
CMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.038
-
CMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0394
-
dCMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.055
-
dCMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.08
-
dCMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.09
-
dCMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.094
-
dCMP
wild-type enzyme
Escherichia coli
0.77
-
dCMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.93
-
UMP
wild-type enzyme
Escherichia coli
0.93
-
UMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1
-
CMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.3
-
CMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.8
-
dCMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.9
-
UMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2.5
-
CMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2.6
-
CMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2.9
-
CMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
3.5
-
dCMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
3.9
-
UMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
5.4
-
UMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
7.3
-
dCMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
7.9
-
UMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
8
-
UMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
11.3
-
UMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
20.2
-
CMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
required
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
24700
-
calculated from amino acid sequence
Escherichia coli K-12
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + CMP
Escherichia coli
-
ADP + CDP
-
-
r
ATP + dCMP
Escherichia coli
-
ADP + dCDP
-
-
r
additional information
Escherichia coli
bacterial CMP kinases are specific for CMP and dCMP, whereas the related eukaryotic NMP kinase phosphorylates CMP and UMP with similar efficiency. Bacterial CMP kinase has a narrower NMP-binding pocket and a hydrogen-bonding network involving the pyrimidine moiety specific for the cytosine nucleobase compared to eukaryotic UMP?CMP kinases
?
-
-
-
Purification (Commentary) (protein specific)
Commentary
Organism
-
Escherichia coli K-12
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
-
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + CMP
-
692282
Escherichia coli
ADP + CDP
-
-
-
r
ATP + CMP
the enzyme is specific for CMP
692282
Escherichia coli K-12
ADP + CDP
-
-
-
?
ATP + dCMP
-
692282
Escherichia coli
ADP + dCDP
-
-
-
r
ATP + dCMP
the enzyme is specific for dCMP
692282
Escherichia coli K-12
ADP + dCDP
-
-
-
?
ATP + UMP
UMP is a poor substrate
692282
Escherichia coli
ADP + UDP
-
-
-
r
ATP + UMP
UMP is a weak substrate
692282
Escherichia coli K-12
ADP + UDP
-
-
-
?
additional information
bacterial CMP kinases are specific for CMP and dCMP, whereas the related eukaryotic NMP kinase phosphorylates CMP and UMP with similar efficiency. Bacterial CMP kinase has a narrower NMP-binding pocket and a hydrogen-bonding network involving the pyrimidine moiety specific for the cytosine nucleobase compared to eukaryotic UMP?CMP kinases
692282
Escherichia coli
?
-
-
-
-
additional information
four key amino acids interacting with the pyrimidine ring of CMP, Ser36, Asp132, Arg110 and Arg188, contribute to the stability, catalysis and substrate specificity of Escherichia coli CMP kinase. D132 is involved in a complicated network of interactions, the strongest being with R110, and also R110 is involved in many interactions, but mainly with D132. Substrate specificities of wild-type and mutant enzymes, overview
692282
Escherichia coli
?
-
-
-
-
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
additional information
-
thermal stability of CMP kinase genetic variants, overview
Escherichia coli
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.013
-
UMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.04
-
dCMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.05
-
dCMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.054
-
UMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.06
-
dCMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.069
-
CMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.12
-
CMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.15
-
dCMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.23
-
CMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.45
-
UMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.57
-
UMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.73
-
dCMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.82
-
UMP
wild-type enzyme
Escherichia coli
0.82
-
UMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.4
-
CMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
4.1
-
CMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
8.3
-
UMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
9.9
-
UMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
14.5
-
dCMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
21.1
-
dCMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
22.4
-
CMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
63
-
CMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
103
-
CMP
wild-type enzyme
Escherichia coli
103
-
CMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
108
-
dCMP
wild-type enzyme
Escherichia coli
108
-
dCMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0033
-
UMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0048
-
UMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0068
-
dCMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0114
-
CMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.052
-
dCMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.053
-
CMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.083
-
UMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.12
-
CMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.3
-
UMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.4
-
dCMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.54
-
CMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.88
-
UMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.04
-
UMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.1
-
dCMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.25
-
UMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.41
-
CMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.88
-
dCMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
4.1
-
dCMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
25.2
-
CMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
230
-
dCMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
589
-
CMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1150
-
dCMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2940
-
CMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0033
-
UMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0048
-
UMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0068
-
dCMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.0114
-
CMP
mutant enzyme R110M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.052
-
dCMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.053
-
CMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.083
-
UMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.12
-
CMP
mutant enzyme R188M, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.3
-
UMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.4
-
dCMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.54
-
CMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
0.88
-
UMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.04
-
UMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.1
-
dCMP
mutant enzyme D132H, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.25
-
UMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.41
-
CMP
mutant enzyme D132A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1.88
-
dCMP
mutant enzyme D132N, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
4.1
-
dCMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
25.2
-
CMP
mutant enzyme S36A, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
230
-
dCMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
589
-
CMP
mutant enzyme D132S, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
1150
-
dCMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
2940
-
CMP
wild type enzyme, in 50 mM Tris-HCl (pH 7.4), 50 mM KCl, 2 mM MgCl2, 1 mM phosphoenolpyruvate, 0.2 mM NADH, at 30°C
Escherichia coli K-12
Other publictions for EC 2.7.4.25
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
712276
Thum
The Rv1712 locus from Mycobact ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Bacteriol.
191
2884-2887
2009
-
-
1
-
-
-
-
3
-
-
3
-
-
160
-
-
1
-
-
-
-
2
6
1
-
-
-
3
-
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
3
-
-
3
-
-
-
-
1
-
-
-
2
6
1
-
-
-
3
-
-
-
-
-
1
1
-
3
3
693707
Caceres
Molecular modeling and dynamic ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Mol. Model.
14
427-434
2008
-
-
1
-
-
-
-
-
-
1
-
2
-
6
-
-
-
-
-
-
-
-
6
1
-
-
-
-
-
-
-
1
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
1
-
2
-
-
-
-
-
-
-
-
6
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
673586
Wang
The role of Ureaplasma nucleos ...
Ureaplasma parvum
FEBS J.
274
1983-1990
2007
-
-
1
-
-
-
-
-
-
1
1
-
-
1
-
-
1
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
-
1
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
676227
Alexandre
Enantioselectivity of human AM ...
Escherichia coli
Nucleic Acids Res.
35
4895-4904
2007
-
-
1
1
7
-
-
23
-
-
-
-
-
1
-
-
1
-
-
-
-
-
3
-
-
-
6
-
-
-
-
-
-
-
-
-
-
1
-
1
7
-
-
-
-
23
-
-
-
-
-
-
-
1
-
-
-
-
3
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
692282
Ofiteru
Structural and functional cons ...
Escherichia coli, Escherichia coli K-12
FEBS J.
274
3363-3373
2007
-
-
2
2
14
1
-
27
-
1
1
3
-
2
-
-
1
-
-
-
1
-
8
-
-
-
1
26
-
-
-
2
-
-
-
-
-
2
2
2
14
1
-
-
-
27
-
1
1
3
-
-
-
1
-
-
1
-
8
-
-
-
1
26
-
-
-
-
-
-
-
-
23
23
675215
Lee
-
Recombinant Escherichia coli-c ...
Escherichia coli
J. Ind. Eng. Chem.
12
757-761
2006
-
1
1
-
-
-
-
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