Cloned (Comment) | Organism |
---|---|
mutant enzyme D93N is expressed in Escherichia coli BL21-CodonPlus(DE3)-RIL cells | Mycobacterium tuberculosis |
Crystallization (Comment) | Organism |
---|---|
mutant enzyme D93N crystallizes to hexagonal plates with 2.0 M ammonium sulfate, 0.1 M Tris-HCl, pH 8.5, and to rods with 2.0 M ammonium sulfate, 2% (w/v) PEG400, 0.1 M HEPES, pH 7.5 | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
D93N | the mutation dramatically decreases protein thermal stability (Tm value of 43.4°C) but is still hexameric and retains full activity. The mutant hexamer is stable and active up to 2.5 M guanidine hydrochloride | Mycobacterium tuberculosis |
General Stability | Organism |
---|---|
guanidinium and many other salts stabilize the hexamer of mutant D93N with a half maximum effect of about 0.1 M, increasing protein thermostability | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Urea | inactivation occurs at very low urea concentrations (0.5 M) | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WJH7 | - |
- |
Mycobacterium tuberculosis H37Rv | P9WJH7 | - |
- |
Purification (Comment) | Organism |
---|---|
Q-Sepharose column chromatography | Mycobacterium tuberculosis |
Renatured (Comment) | Organism |
---|---|
the wild type enzyme unfolds and inactivates simultaneously in the presence of 4 M guanidine hydrochloride as with 4 M urea as denaturant. Refolding occurs at much lower guanidine hydrochloride concentrations (0.5-1 M) | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 8-bromoinosine 5'-diphosphate | - |
Mycobacterium tuberculosis | ADP + 8-bromoinosine 5'-triphosphate | - |
? | |
ATP + 8-bromoinosine 5'-diphosphate | - |
Mycobacterium tuberculosis H37Rv | ADP + 8-bromoinosine 5'-triphosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | x-ray crystallography | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
NDPK | - |
Mycobacterium tuberculosis |
nucleoside diphosphate kinase | - |
Mycobacterium tuberculosis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
76 | - |
the wild type enzyme has a temperature of half denaturation of 76°C | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.159 | - |
8-bromoinosine 5'-diphosphate | wild type enzyme, at pH 7.5 and 22°C | Mycobacterium tuberculosis | |
0.23 | - |
8-bromoinosine 5'-diphosphate | mutant enzyme D93N, at pH 7.5 and 22°C | Mycobacterium tuberculosis | |
232 | - |
ATP | mutant enzyme D93N, at pH 7.5 and 22°C | Mycobacterium tuberculosis | |
264 | - |
ATP | wild type enzyme, at pH 7.5 and 22°C | Mycobacterium tuberculosis |