Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | strong stimulation of glutaminase subunit TrpG activity by the associated synthase subunit TrpE within the glutamine amidotransferase, GATase, enzyme complex | Thermotoga maritima |
Cloned (Comment) | Organism |
---|---|
gene trpG, expression of His-tagged wild-type TrpG and mutant TrpG variants T129F and T129A in Escherichia coli strain BL21(DE3)Rosetta | Thermotoga maritima |
Protein Variants | Comment | Organism |
---|---|---|
L126G | site-directed mutagenesis, the mutant shows constitutive activity unlike the wild-type enzyme and does not require presence of TrpE | Thermotoga maritima |
L126G/V127Y/T129Y/Y131V | site-directed mutagenesis, the mutant shows constitutive activity unlike the wild-type enzyme and does not require presence of TrpE | Thermotoga maritima |
T129Y | site-directed mutagenesis, the mutant shows constitutive activity unlike the wild-type enzyme and does not require presence of TrpE | Thermotoga maritima |
V127Y | site-directed mutagenesis, the mutant shows no constitutive activity like the wild-type enzyme and requires presence of TrpE | Thermotoga maritima |
Y131V | site-directed mutagenesis, the mutant shows no constitutive activity like the wild-type enzyme and requires presence of TrpE | Thermotoga maritima |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | glutaminase activity steady-state enzyme kinetics of TrpG and the AS complex, generated by mixing the recombinant TrpE and TrpGD subunits | Thermotoga maritima |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamine + H2O | Thermotoga maritima | - |
L-glutamate + NH3 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga maritima | - |
gene trpG | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged TrpG, and mutant TrpG variants T129F and T129A from Escherichia coli strain BL21(DE3)Rosetta by heat treatment for 20 min at 60°C, nickel affinity chromatography, and anion exchange chromatography | Thermotoga maritima |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamine + H2O | - |
Thermotoga maritima | L-glutamate + NH3 | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the glutaminase subunit TrpG, which adopts the triad G-type amidotransferase fold, and the synthase subunit TrpE, which adopts a complicated alpha/beta folding pattern, assemble to heterotetrameric (TrpE/TrpG)2-complexes. The sequence stretch L126-V127-A128-T129-R130-Y131 is localized between the catalytic triad Cys83-His175-Glu177 and the associated TrpE subunit, homology modeling based on the structure of Sulfolobus solfataricus TrpG | Thermotoga maritima |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Thermotoga maritima |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Thermotoga maritima |
Organism | Comment | Expression |
---|---|---|
Thermotoga maritima | wild-type TrpG shows no constitutive activity and needs to be induced by TrpE | up |