Protein Variants | Comment | Organism |
---|---|---|
K258A | about 2600fold decrease in kcat/Km for glutamine dependent cyclase reaction, basal and stimulated glutaminase functions are not altered. The stoichiometry of reaction glutamine: 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide turnover is 43:1 compared to wild-type value 1:1 | Saccharomyces cerevisiae |
K258R | about 20fold decrease in kcat/Km for glutamine dependent cyclase reaction,basal and stimulated glutaminase functions are not altered. The stoichiometry of reaction glutamine: 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide turnover is 43:1 compared to wild-type value 3:1 | Saccharomyces cerevisiae |
K360A | synthase activity is similar to wild type. The stoichiometry of the reaction is 3:1 indicating some degree of uncoupling due to loss of ammonia | Saccharomyces cerevisiae |
K360R | synthase activity is similar to wild type. The mutant retains both structural and functional integrity of the enzyme in the case of glutaminase functions | Saccharomyces cerevisiae |
R239A | a 1000fold reduction in kcat/Km is observed in the glutamine dependent synthase reaction. The stoichiometry of reaction glutamine: 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide turnover is 43:1 compared to wild-type value 122:1 | Saccharomyces cerevisiae |
R239H | a 1000fold reduction in kcat/Km is observed in the glutamine dependent synthase reaction. The stoichiometry of reaction glutamine: 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide turnover is 43:1 compared to wild-type value 154:1 | Saccharomyces cerevisiae |
R239K | a 1000fold reduction in kcat/Km is observed in the glutamine dependent synthase reaction. The stoichiometry of reaction glutamine: 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide turnover is 43:1 compared to wild-type value 40:1 | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
albizziin | mixed-type inhibitor versus 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide, competitive versus glutamine | Saccharomyces cerevisiae | |
rPRFAR-1 | competitive inhibitor versus 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide inhibitor, mixed-type inhibitor versus glutamine | Saccharomyces cerevisiae | |
rPRFAR-2 | competitive versus 5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0016 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239K | Saccharomyces cerevisiae | |
0.0018 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K360A | Saccharomyces cerevisiae | |
0.002 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K258R | Saccharomyces cerevisiae | |
0.0023 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K360R | Saccharomyces cerevisiae | |
0.003 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239A | Saccharomyces cerevisiae | |
0.005 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, wild-type enzyme | Saccharomyces cerevisiae | |
0.0079 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239H | Saccharomyces cerevisiae | |
0.046 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K258R | Saccharomyces cerevisiae | |
0.053 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239A | Saccharomyces cerevisiae | |
0.055 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, wild-type enzyme | Saccharomyces cerevisiae | |
0.065 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K360A | Saccharomyces cerevisiae | |
0.072 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K360R | Saccharomyces cerevisiae | |
0.075 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239H | Saccharomyces cerevisiae | |
0.08 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239K | Saccharomyces cerevisiae | |
0.098 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K258A | Saccharomyces cerevisiae | |
0.139 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K258A | Saccharomyces cerevisiae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P33734 | - |
- |
Saccharomyces cerevisiae ATCC 204508 | P33734 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine | 4900fold upregulation of glutamine hydrolysis in the presence of an acceptor substrate. Transfer of ammonia from the glutaminase site occurs through the (beta/alpha)(8) core of the protein. The conserved K258 residue is key to productive binding and the overall stoichiometry of the reaction. The binding of the ribulosyl phosphate portion of the substrate appears to be transduced through reorientation of K258 resulting in a conformational switch at the base of the (beta/alpha)(8) core that enables the passage of ammonia through the core of the protein | Saccharomyces cerevisiae | 5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate | - |
? | |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + L-glutamine | 4900fold upregulation of glutamine hydrolysis in the presence of an acceptor substrate. Transfer of ammonia from the glutaminase site occurs through the (beta/alpha)(8) core of the protein. The conserved K258 residue is key to productive binding and the overall stoichiometry of the reaction. The binding of the ribulosyl phosphate portion of the substrate appears to be transduced through reorientation of K258 resulting in a conformational switch at the base of the (beta/alpha)(8) core that enables the passage of ammonia through the core of the protein | Saccharomyces cerevisiae ATCC 204508 | 5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + L-glutamate | - |
? | |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + NH4+ | - |
Saccharomyces cerevisiae | 5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + H2O | - |
? | |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + NH4+ | - |
Saccharomyces cerevisiae ATCC 204508 | 5-amino-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide + D-erythro-1-(imidazol-4-yl)glycerol 3-phosphate + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
IGP synthase | - |
Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K258A | Saccharomyces cerevisiae | |
0.126 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K258R | Saccharomyces cerevisiae | |
0.15 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239A | Saccharomyces cerevisiae | |
0.21 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239H | Saccharomyces cerevisiae | |
0.24 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K360A | Saccharomyces cerevisiae | |
0.29 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K360R | Saccharomyces cerevisiae | |
0.3 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239K | Saccharomyces cerevisiae | |
0.31 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K360R | Saccharomyces cerevisiae | |
0.7 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K360A | Saccharomyces cerevisiae | |
0.845 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, wild-type enzyme | Saccharomyces cerevisiae | |
3.9 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239H | Saccharomyces cerevisiae | |
4.3 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239A | Saccharomyces cerevisiae | |
5 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K258R | Saccharomyces cerevisiae | |
5.4 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, wild-type enzyme | Saccharomyces cerevisiae | |
5.4 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K258A | Saccharomyces cerevisiae | |
8.7 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239K | Saccharomyces cerevisiae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.039 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K258A | Saccharomyces cerevisiae | |
0.12 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K258R | Saccharomyces cerevisiae | |
0.46 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K258A | Saccharomyces cerevisiae | |
0.5 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239H | Saccharomyces cerevisiae | |
1.4 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239A | Saccharomyces cerevisiae | |
2.8 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239A | Saccharomyces cerevisiae | |
2.9 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239H | Saccharomyces cerevisiae | |
3.8 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme R239K | Saccharomyces cerevisiae | |
4.3 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K360R | Saccharomyces cerevisiae | |
5.5 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme R239K | Saccharomyces cerevisiae | |
11 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, mutant enzyme K360A | Saccharomyces cerevisiae | |
60 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K258R | Saccharomyces cerevisiae | |
110 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K360R | Saccharomyces cerevisiae | |
130 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, mutant enzyme K360A | Saccharomyces cerevisiae | |
150 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: NH4+, wild-type enzyme | Saccharomyces cerevisiae | |
1200 | - |
5-[(5-phospho-1-deoxy-D-ribulos-1-ylamino)methylideneamino]-1-(5-phospho-beta-D-ribosyl)imidazole-4-carboxamide | 30°C, pH 7.0, cosubstrate: L-glutamine, wild-type enzyme | Saccharomyces cerevisiae |