Application | Comment | Organism |
---|---|---|
pharmacology | enzyme is a potential drug target in the treatment of parasite caused disease | Trypanosoma cruzi |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli | Trypanosoma cruzi |
Protein Variants | Comment | Organism |
---|---|---|
G69S | reduced activity compared to the wild-type enzyme | Trypanosoma cruzi |
L67M | highly reduced activity with hypoxanthine compared to the wild-type enzyme, mutant is not active with guanine | Trypanosoma cruzi |
additional information | saturation mutagenesis and complement selection for investigation of functional roles of residues Leu67 and Gly69, sequencing of 70 clones and identification of 30 different mutations, several mutants of L67 or G69 support bacterial growth on minimal medium, but only L67M and G69S can be expressed and purified from overexpressing bacteria, overview | Trypanosoma cruzi |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0086 | - |
hypoxanthine | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
0.012 | - |
guanine | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
0.0192 | - |
hypoxanthine | pH 7.5, 37°C, recombinant mutant L67M | Trypanosoma cruzi | |
0.022 | - |
guanine | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
0.027 | - |
IMP | pH 7.5, 37°C, recombinant wild-type enzyme and mutant L67M | Trypanosoma cruzi | |
0.032 | - |
5-phospho-alpha-D-ribose 1-diphosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
0.0339 | - |
hypoxanthine | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
0.035 | - |
diphosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
0.08 | - |
IMP | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
0.083 | - |
5-phospho-alpha-D-ribose 1-diphosphate | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
0.132 | - |
5-phospho-alpha-D-ribose 1-diphosphate | pH 7.5, 37°C, recombinant mutant L67M | Trypanosoma cruzi | |
0.135 | - |
diphosphate | pH 7.5, 37°C, recombinant mutant L67M | Trypanosoma cruzi | |
0.214 | - |
diphosphate | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | at 12 mM | Trypanosoma cruzi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GMP + diphosphate | Trypanosoma cruzi | - |
guanine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
r | |
IMP + diphosphate | Trypanosoma cruzi | - |
hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma cruzi | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli by GMP affinity and cation exchange chromatography | Trypanosoma cruzi |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
IMP + diphosphate = hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | reaction mechanism, function and influence of invariant active site loop I residues on enzyme efficiency in forward and reverse reaction | Trypanosoma cruzi |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Trypanosoma cruzi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GMP + diphosphate | - |
Trypanosoma cruzi | guanine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
r | |
IMP + diphosphate | - |
Trypanosoma cruzi | hypoxanthine + 5-phospho-alpha-D-ribose 1-diphosphate | - |
r | |
additional information | L67 side chain participates in hydrophobic interactions between dimer subunits important for the proper positioning of main chain atoms that influence enzyme chemistry and the binding of 5-phospho-alpha-D-ribose 1-diphosphate, diphosphate, and hypoxanthine, residue 69 needs to be small like Gly, which is found there, to avoid sterical interference with binding of 5-phospho-alpha-D-ribose 1-diphosphate and diphosphate as well as the positioning of a metal ion | Trypanosoma cruzi | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
HPRT | - |
Trypanosoma cruzi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Trypanosoma cruzi |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
IMP | pH 7.5, 37°C, recombinant mutant L67M | Trypanosoma cruzi | |
0.12 | - |
diphosphate | pH 7.5, 37°C, recombinant mutant L67M | Trypanosoma cruzi | |
0.19 | - |
IMP | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
0.3 | - |
hypoxanthine | pH 7.5, 37°C, recombinant mutant L67M | Trypanosoma cruzi | |
0.31 | - |
diphosphate | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
0.47 | - |
IMP | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
0.48 | - |
diphosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
4.7 | - |
5-phospho-alpha-D-ribose 1-diphosphate | pH 7.5, 37°C, recombinant mutant L67M | Trypanosoma cruzi | |
20 | - |
5-phospho-alpha-D-ribose 1-diphosphate | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
22 | - |
hypoxanthine | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
22.1 | - |
guanine | pH 7.5, 37°C, recombinant mutant G69S | Trypanosoma cruzi | |
22.9 | - |
hypoxanthine | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
23.2 | - |
5-phospho-alpha-D-ribose 1-diphosphate | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi | |
32.9 | - |
guanine | pH 7.5, 37°C, recombinant wild-type enzyme | Trypanosoma cruzi |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Trypanosoma cruzi |