Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Escherichia coli K-12 |
expression of wild-type and mutant enzymes | Escherichia coli |
Crystallization (Comment) | 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 |
Protein Variants | Comment | 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 | 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 Maximum [mM] | Substrate | Comment | 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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
24700 | - |
calculated from amino acid sequence | Escherichia coli K-12 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A6I0 | - |
- |
Escherichia coli K-12 | P0A6I0 | strain CJ236 | - |
Purification (Comment) | Organism |
---|---|
- |
Escherichia coli K-12 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + CMP | - |
Escherichia coli | ADP + CDP | - |
r | |
ATP + CMP | the enzyme is specific for CMP | Escherichia coli K-12 | ADP + CDP | - |
? | |
ATP + dCMP | - |
Escherichia coli | ADP + dCDP | - |
r | |
ATP + dCMP | the enzyme is specific for dCMP | Escherichia coli K-12 | ADP + dCDP | - |
? | |
ATP + UMP | UMP is a poor substrate | Escherichia coli | ADP + UDP | - |
r | |
ATP + UMP | UMP is a weak substrate | 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 | 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 | Escherichia coli | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CMP kinase | - |
Escherichia coli |
CMP kinase | - |
Escherichia coli K-12 |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
thermal stability of CMP kinase genetic variants, overview | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | 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 | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli | |
ATP | - |
Escherichia coli K-12 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | 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 |