BRENDA - Enzyme Database

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)
EC Number
Commentary
Organism
2.7.4.25
expressed in Escherichia coli BL21(DE3) cells
Escherichia coli K-12
2.7.4.25
expression of wild-type and mutant enzymes
Escherichia coli
Crystallization (Commentary)
EC Number
Crystallization
Organism
2.7.4.25
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
2.7.4.25
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
EC Number
Amino acid exchange
Commentary
Organism
2.7.4.25
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
2.7.4.25
D132A
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
2.7.4.25
D132H
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
2.7.4.25
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
2.7.4.25
D132N
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
2.7.4.25
D132N
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
2.7.4.25
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli K-12
2.7.4.25
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
2.7.4.25
R110M
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
EC Number
General Stability
Organism
2.7.4.25
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]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.4.25
additional information
-
additional information
kinetics and substrate specificities of wild-type and mutant enzymes, overview
Escherichia coli
2.7.4.25
0.035
-
CMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
0.094
-
dCMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
0.93
-
UMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.4.25
Mg2+
required
Escherichia coli
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.4.25
24700
-
calculated from amino acid sequence
Escherichia coli K-12
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.4.25
ATP + CMP
Escherichia coli
-
ADP + CDP
-
-
r
2.7.4.25
ATP + dCMP
Escherichia coli
-
ADP + dCDP
-
-
r
2.7.4.25
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
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.4.25
Escherichia coli
P0A6I0
-
-
2.7.4.25
Escherichia coli K-12
P0A6I0
strain CJ236
-
Purification (Commentary)
EC Number
Commentary
Organism
2.7.4.25
-
Escherichia coli K-12
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.4.25
additional information
-
-
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.4.25
ATP + CMP
-
692282
Escherichia coli
ADP + CDP
-
-
-
r
2.7.4.25
ATP + CMP
the enzyme is specific for CMP
692282
Escherichia coli K-12
ADP + CDP
-
-
-
?
2.7.4.25
ATP + dCMP
-
692282
Escherichia coli
ADP + dCDP
-
-
-
r
2.7.4.25
ATP + dCMP
the enzyme is specific for dCMP
692282
Escherichia coli K-12
ADP + dCDP
-
-
-
?
2.7.4.25
ATP + UMP
UMP is a poor substrate
692282
Escherichia coli
ADP + UDP
-
-
-
r
2.7.4.25
ATP + UMP
UMP is a weak substrate
692282
Escherichia coli K-12
ADP + UDP
-
-
-
?
2.7.4.25
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
?
-
-
-
-
2.7.4.25
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]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.4.25
additional information
-
thermal stability of CMP kinase genetic variants, overview
Escherichia coli
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
0.82
-
UMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
103
-
CMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
108
-
dCMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
EC Number
Cofactor
Commentary
Organism
Structure
2.7.4.25
ATP
-
Escherichia coli K-12
2.7.4.25
ATP
-
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.4.25
expressed in Escherichia coli BL21(DE3) cells
Escherichia coli K-12
2.7.4.25
expression of wild-type and mutant enzymes
Escherichia coli
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
2.7.4.25
ATP
-
Escherichia coli
2.7.4.25
ATP
-
Escherichia coli K-12
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
2.7.4.25
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
2.7.4.25
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)
EC Number
Amino acid exchange
Commentary
Organism
2.7.4.25
D132A
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
2.7.4.25
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
2.7.4.25
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
2.7.4.25
D132H
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
2.7.4.25
D132N
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
2.7.4.25
D132N
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
2.7.4.25
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli
2.7.4.25
D132S
the substitution results in a moderate decrease in stability without significant changes in Km value for CMP and dCMP
Escherichia coli K-12
2.7.4.25
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
2.7.4.25
R110M
the mutant shows reduced activity compared to the wild type enzyme
Escherichia coli K-12
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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)
EC Number
General Stability
Organism
2.7.4.25
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)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7.4.25
additional information
-
additional information
kinetics and substrate specificities of wild-type and mutant enzymes, overview
Escherichia coli
2.7.4.25
0.035
-
CMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
0.094
-
dCMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
0.93
-
UMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.7.4.25
Mg2+
required
Escherichia coli
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.7.4.25
24700
-
calculated from amino acid sequence
Escherichia coli K-12
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.4.25
ATP + CMP
Escherichia coli
-
ADP + CDP
-
-
r
2.7.4.25
ATP + dCMP
Escherichia coli
-
ADP + dCDP
-
-
r
2.7.4.25
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)
EC Number
Commentary
Organism
2.7.4.25
-
Escherichia coli K-12
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.7.4.25
additional information
-
-
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.4.25
ATP + CMP
-
692282
Escherichia coli
ADP + CDP
-
-
-
r
2.7.4.25
ATP + CMP
the enzyme is specific for CMP
692282
Escherichia coli K-12
ADP + CDP
-
-
-
?
2.7.4.25
ATP + dCMP
-
692282
Escherichia coli
ADP + dCDP
-
-
-
r
2.7.4.25
ATP + dCMP
the enzyme is specific for dCMP
692282
Escherichia coli K-12
ADP + dCDP
-
-
-
?
2.7.4.25
ATP + UMP
UMP is a poor substrate
692282
Escherichia coli
ADP + UDP
-
-
-
r
2.7.4.25
ATP + UMP
UMP is a weak substrate
692282
Escherichia coli K-12
ADP + UDP
-
-
-
?
2.7.4.25
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
?
-
-
-
-
2.7.4.25
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)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
2.7.4.25
additional information
-
thermal stability of CMP kinase genetic variants, overview
Escherichia coli
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
0.82
-
UMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
103
-
CMP
wild-type enzyme
Escherichia coli
2.7.4.25
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
2.7.4.25
108
-
dCMP
wild-type enzyme
Escherichia coli
2.7.4.25
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]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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
2.7.4.25
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