Crystallization (Comment) | Organism |
---|---|
GlnRS-tRNAGln complex, 6.6 mg/ml protein in 10 mM PIPES, pH 7.5, 10 mM MgCl2, and 1.8-5.4 mM tRNA. The tRNA/analog solution is then mixed with equal volumes of a 6.3 mg/ml solution of GlnRS, containing 5mM PIPES, pH 7.0, and 5 mM 2-mercaptoethanol, X-ray diffraction structure determination and analysis at 2.6 A resolution | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
additional information | the engineered mutant hybrid C229R Gln-RS, EC 6.1.1.18, shows activity with L-glutamine or L-glutamate and tRNAGln like the nondiscriminating enzyme, EC 6.1.1.24. Introduction of 22 amino acid replacements and one deletion, including substitution of the entire primary binding site and two surface loops adjacent to the region disrupted in the mutant C229R, improves the capacity of the mutant enzyme to synthesize misacylated Glu-tRNAGln by 16000fold, overview | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of the mutant enzyme compared to wild-type GlnRS, EC 6.1.1.18, overview | Escherichia coli | |
0.21 | - |
L-glutamine | mutant C229R GlnRS, with tRNAGln | Escherichia coli | |
240 | - |
L-glutamate | mutant C229R GlnRS, with tRNAGln | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate + tRNAGln | Escherichia coli | - |
AMP + diphosphate + L-glutamyl-tRNAGln | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P00962 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate + tRNAGln | - |
Escherichia coli | AMP + diphosphate + L-glutamyl-tRNAGln | - |
? | |
ATP + L-glutamate + tRNAGln | activity also with Gln-RS, EC 6.1.1.18, mutant C229R | Escherichia coli | AMP + diphosphate + L-glutamyl-tRNAGln | - |
? | |
ATP + L-glutamine + tRNAGln | - |
Escherichia coli | AMP + diphosphate + glutaminyl-tRNAGln | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00041 | - |
L-glutamate | mutant C229R GlnRS, with tRNAGln | Escherichia coli | |
0.0025 | - |
L-glutamine | mutant C229R GlnRS, with tRNAGln | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |