Activating Compound | Comment | Organism | Structure |
---|---|---|---|
5-fluoro-UTP | below 0.15 mM 5-fluoro-UTP increases the activity of the enzyme by 80% | Mycobacterium tuberculosis | |
GMP | much weaker activator (only 30% increase in activity at 2.5 mM) | Mycobacterium tuberculosis | |
GMP-4-nitrophenol | much weaker activator (only 50% increase in activity at 1 mM) | Mycobacterium tuberculosis | |
GTP | GTP increases the reaction rate by a factor of 3.5, with half-saturation at 0.1 mM | Mycobacterium tuberculosis | |
additional information | GMP-PCP and 8-bromo-GTP are ineffective as activators | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)/pDIA17 cells | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
using sodium/potassium tartrate (1.2 M) at pH 7.4 and 10 mM GTP | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
D113A | the mutant shows an increased Km value for UMP compared to the wild type enzyme | Mycobacterium tuberculosis |
F81W | the mutant shows an increased Km value for UMP compared to the wild type enzyme | Mycobacterium tuberculosis |
F81W/S96A | the mutant shows a decreased Km value for UMP compared to the wild type enzyme | Mycobacterium tuberculosis |
P139A | the mutant shows a decreased Km value for UMP compared to the wild type enzyme | Mycobacterium tuberculosis |
P139H | the mutant shows a decreased Km value for UMP compared to the wild type enzyme | Mycobacterium tuberculosis |
P139W | the mutant shows a decreased Km value for UMP compared to the wild type enzyme | Mycobacterium tuberculosis |
R150A | the mutant shows a decreased Km value for UMP compared to the wild type enzyme | Mycobacterium tuberculosis |
R82H | the mutant shows an increased Km value for UMP compared to the wild type enzyme and 0.7% of the wild-type specific activity | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5-bromo-UMP | weak competitive inhibitor | Mycobacterium tuberculosis | |
5-fluoro-UTP | at 0.6mM 5-fluoro-UTP, the enzyme activity is decreased by 40% | Mycobacterium tuberculosis | |
5-iodo-UTP | strongest inhibitor | Mycobacterium tuberculosis | |
dTTP | - |
Mycobacterium tuberculosis | |
dUMP | weak competitive inhibitor | Mycobacterium tuberculosis | |
dUTP | - |
Mycobacterium tuberculosis | |
UTP | - |
Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0023 | - |
UMP | mutant enzyme P139H, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.0034 | - |
UMP | mutant enzyme F81W/S96A, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.0061 | - |
UMP | mutant enzyme P139W, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.0066 | - |
UMP | mutant enzyme P139A, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.014 | - |
UMP | mutant enzyme R150A, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.0177 | - |
UMP | wild type enzyme, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.0639 | - |
UMP | mutant enzyme F81W, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.0891 | - |
UMP | mutant enzyme R82H, at pH 7.4 and 30°C | Mycobacterium tuberculosis | |
0.2 | - |
UMP | mutant enzyme D113A, at pH 7.4 and 30°C | Mycobacterium tuberculosis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Mycobacterium tuberculosis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
173200 | - |
sedimentation equilibrium ultracentrifugation | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WHK5 | - |
- |
Mycobacterium tuberculosis H37Rv | P9WHK5 | - |
- |
Purification (Comment) | Organism |
---|---|
TALON resin column chromatography and Superdex 200 gel filtration | Mycobacterium tuberculosis |
Storage Stability | Organism |
---|---|
4°C, in 20 mM phosphate (pH 7.0) and 100 mM NaCl, about 2 months, no apparent loss of activity | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + UMP | - |
Mycobacterium tuberculosis | ADP + UDP | - |
? | |
ATP + UMP | - |
Mycobacterium tuberculosis H37Rv | ADP + UDP | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | - |
Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
Rv2883c | - |
Mycobacterium tuberculosis |
UMPK | - |
Mycobacterium tuberculosis |
uridine monophosphate kinase | - |
Mycobacterium tuberculosis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | 75 | the enzyme shows Tm values of 60°C in the absence of nucleotides and 65°C and 75°C with 1 mM GTP and UTP, respectively | Mycobacterium tuberculosis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
5-bromo-UMP | Ki above 1 mM, wild type enzyme, at pH 7.4 and 30°C | Mycobacterium tuberculosis |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.02 | - |
wild type enzyme, at pH 7.4 and 30°C | Mycobacterium tuberculosis | 5-iodo-UTP | |
0.1 | - |
wild type enzyme, at pH 7.4 and 30°C | Mycobacterium tuberculosis | UTP | |
0.24 | - |
wild type enzyme, at pH 7.4 and 30°C | Mycobacterium tuberculosis | dUTP | |
0.4 | - |
wild type enzyme, at pH 7.4 and 30°C | Mycobacterium tuberculosis | dTTP |