Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
in complex with substrates CMP, dCMP, ara-CMP and 2,3-dideoxy-CMP, hanging drop vapor diffusion method, using ammonium sulfate in a 50 mM Tris-HCl buffer (pH 7.4), at 20°C | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D185A | the mutant shows severely decreased CMP and dCMP phosphoryation activity compared to the wild type enzyme | Escherichia coli |
R181M | the mutant shows severely decreased CMP and dCMP phosphoryation activity compared to the wild type enzyme | Escherichia coli |
S101A | the mutation reduces CMP phosphorylation only moderately, but dramatically reduces dCMP phosphorylation | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.035 | - |
CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli | |
0.08 | - |
CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
0.094 | - |
dCMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli | |
0.15 | - |
2',3'-dideoxy-CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli | |
0.19 | - |
CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
0.19 | - |
dCMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
0.24 | - |
dCMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli | |
0.24 | - |
dCMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
0.46 | - |
2',3'-dideoxy-CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli | |
0.47 | - |
CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli | |
0.47 | - |
ara-CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
0.53 | - |
ara-CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli | |
0.54 | - |
2',3'-dideoxy-CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
0.65 | - |
2',3'-dideoxy-CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
0.79 | - |
ara-CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
1 | - |
ara-CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required for activity | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + CMP | Escherichia coli | - |
ADP + CDP | - |
? | |
ATP + dCMP | Escherichia coli | Escherichia coli CMPK phosphorylates dCMP nearly as well as it does CMP | ADP + dCDP | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | A0A140N8S7 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2',3'-dideoxy-CMP | 2,3-dideoxy-CMP is a poor substrate | Escherichia coli | ADP + ? | - |
? | |
ATP + ara-CMP | ara-CMP is a poor substrate | Escherichia coli | ADP + ara-CDP | - |
? | |
ATP + CMP | - |
Escherichia coli | ADP + CDP | - |
? | |
ATP + dCMP | Escherichia coli CMPK phosphorylates dCMP nearly as well as it does CMP | Escherichia coli | ADP + dCDP | - |
? | |
ATP + UMP | bacterial CMP kinases phosphorylate UMP with very low rates | Escherichia coli | ADP + UDP | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CMP kinase | - |
Escherichia coli |
CMPK | - |
Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
52 | - |
the melting temperature of the wild type enzyme is at 52°C | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0083 | - |
2',3'-dideoxy-CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli | |
0.047 | - |
2',3'-dideoxy-CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli | |
0.071 | - |
dCMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli | |
0.08 | - |
CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
0.085 | - |
ara-CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli | |
0.12 | - |
2',3'-dideoxy-CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
0.19 | - |
dCMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
0.26 | - |
CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli | |
0.45 | - |
dCMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
0.47 | - |
ara-CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
0.65 | - |
2',3'-dideoxy-CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli | |
1.36 | - |
ara-CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
1.38 | - |
CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli | |
56 | - |
ara-CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli | |
103 | - |
CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli | |
109 | - |
dCMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.083 | - |
ara-CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli K-12 | |
0.3 | - |
dCMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli K-12 | |
0.54 | - |
CMP | mutant enzyme D185A, at 30°C and pH 7.4 | Escherichia coli K-12 | |
1.7 | - |
ara-CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli K-12 | |
1.9 | - |
dCMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli K-12 | |
6.1 | - |
dCMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli K-12 | |
7.4 | - |
CMP | mutant enzyme R181M, at 30°C and pH 7.4 | Escherichia coli K-12 | |
7.5 | - |
ara-CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli K-12 | |
105 | - |
ara-CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli K-12 | |
697 | - |
CMP | mutant enzyme S101A, at 30°C and pH 7.4 | Escherichia coli K-12 | |
1160 | - |
dCMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli K-12 | |
2940 | - |
CMP | wild type enzyme, at 30°C and pH 7.4 | Escherichia coli K-12 |