Cloned (Comment) | Organism |
---|---|
functional expression of wild-type enzyme and mutants in an enzyme-deficient Escherichia coli strain | Leishmania donovani |
Protein Variants | Comment | Organism |
---|---|---|
additional information | wild-type enzyme complements enzyme deficiency of the bacterial enzyme in Escherichia coli | Leishmania donovani |
S95A | site-directed mutagenesis, dramatic reduction of catalytic activity, weak complementation of bacterial enzyme deficient strain | Leishmania donovani |
S95C | site-directed mutagenesis, 2-3fold reduction of kcat, weak complementation of bacterial enzyme deficient strain | Leishmania donovani |
S95E | site-directed mutagenesis, dramatic reduction of catalytic activity, no complementation of bacterial enzyme deficient strain | Leishmania donovani |
S95T | site-directed mutagenesis, 2-3fold reduction of kcat, complementation of bacterial enzyme deficient strain | Leishmania donovani |
Y96F | site-directed mutagenesis, dramatic reduction of catalytic activity, no complementation of bacterial enzyme deficient strain, 4-5fold decrease of Km value for 5-phosphoribosyl 1-diphosphate | Leishmania donovani |
Y96V | site-directed mutagenesis, dramatic reduction of catalytic activity, no complementation of bacterial enzyme deficient strain, 4-5fold decrease of Km value for 5-phosphoribosyl 1-diphosphate | Leishmania donovani |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
diethyl dicarbonate | alkylation of Arg155, complete inactivation at pH 9.0, pH dependent | Leishmania donovani | |
additional information | no effect of iodoacetate, phenylglyoxal, p-chloromercuribenzoate, acetic anhydride, ethyl dimethylaminopropylcarbodiimide/ammonium acetate, and diisopropyl fluorophosphate | Leishmania donovani | |
Tetranitromethane | complete inactivation at pH 9.0, pH dependent, modifies Tyr96 in the active site | Leishmania donovani |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Leishmania donovani | |
additional information | - |
additional information | Km values of mutant enzymes for hypoxanthine, guanine, diphosphate, 5-phosphoribosyl 1-diphosphate, inosine monophosphate | Leishmania donovani | |
0.005 | - |
guanine | wild-type enzyme | Leishmania donovani | |
0.0061 | - |
hypoxanthine | wild-type enzyme | Leishmania donovani | |
0.09 | - |
inosine monophosphate | wild-type enzyme | Leishmania donovani | |
0.103 | - |
diphosphate | wild-type enzyme | Leishmania donovani | |
0.134 | - |
5-phospho-alpha-D-ribose 1-diphosphate | wild-type enzyme | Leishmania donovani |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Leishmania donovani |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
guanine + 5-phospho-alpha-D-ribose 1-diphosphate | Leishmania donovani | - |
GMP + diphosphate | - |
? | |
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | Leishmania donovani | - |
IMP + diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Leishmania donovani | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant from Escherichia coli | Leishmania donovani |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
IMP + diphosphate = hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | essential role of Ser95-Tyr96 dyade in catalysis | Leishmania donovani |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
kinetics | Leishmania donovani |
Storage Stability | Organism |
---|---|
-70°C, wild-type enzyme and mutants, except mutant S95C, stable up to 6 months | Leishmania donovani |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
guanine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
Leishmania donovani | GMP + diphosphate | - |
? | |
guanine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
Leishmania donovani | GMP + diphosphate | - |
r | |
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
Leishmania donovani | IMP + diphosphate | - |
? | |
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
Leishmania donovani | IMP + diphosphate | - |
r | |
additional information | kinetic study | Leishmania donovani | ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Leishmania donovani |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8.54 | - |
hypoxanthine | wild-type enzyme | Leishmania donovani | |
36.3 | - |
5-phospho-alpha-D-ribose 1-diphosphate | wild-type enzyme | Leishmania donovani | |
41.3 | - |
guanine | wild-type enzyme | Leishmania donovani | |
77 | - |
inosine monophosphate | wild-type enzyme | Leishmania donovani | |
90 | - |
diphosphate | wild-type enzyme | Leishmania donovani |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Leishmania donovani |