Cloned (Comment) | Organism |
---|---|
expression of C-terminally and N-terminally His6-tagged ND-GluRS in Escherichia coli strain BL21 | Thermosynechococcus vestitus |
Crystallization (Comment) | Organism |
---|---|
GluRS in complex with glutamate, hanging drop vapor diffusion at 20°C, 0.003 ml protein solution containing 3 mg/ml protein in 20 mM HEPES, pH 7.9, 20 mM NaCl, 10 mM DTT, 0.25 mM zinc acetate and 0.25 mM MgCl2, is mixed with 0.003 ml reservoir solution containing 740 mM sodium citrate, 140 mM citric acid, pH 5.8, and 10 mM DTT, 1-2 weeks, cryoprotection using 25% v/v of a 50% w/v trehalose solution, X-ray diffraction structure determination and analysis at 2.45 A resolution | Thermosynechococcus vestitus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Thermosynechococcus vestitus | |
0.00079 | - |
tRNAGlu | - |
Thermosynechococcus vestitus | |
0.0037 | - |
tRNAGln | - |
Thermosynechococcus vestitus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Thermosynechococcus vestitus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate + tRNAGln | Thermosynechococcus vestitus | tRNAGln is initially mischarged with glutamate by a non-discriminating glutamyl-tRNA synthetase | AMP + diphosphate + L-glutamyl-tRNAGln | - |
? | |
ATP + L-glutamate + tRNAGlu | Thermosynechococcus vestitus | - |
AMP + diphosphate + L-glutamyl-tRNAGlu | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermosynechococcus vestitus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally and N-terminally His6-tagged ND-GluRS from Escherichia coli strain BL21 by nickel affinity and anion exchange chromatography, and gel filtration to homogeneity | Thermosynechococcus vestitus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + L-glutamate + tRNAGlx = AMP + diphosphate + L-glutamyl-tRNAGlx | active site structure and substrate recognition | Thermosynechococcus vestitus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate + tRNAGln | tRNAGln is initially mischarged with glutamate by a non-discriminating glutamyl-tRNA synthetase | Thermosynechococcus vestitus | AMP + diphosphate + L-glutamyl-tRNAGln | - |
? | |
ATP + L-glutamate + tRNAGlu | - |
Thermosynechococcus vestitus | AMP + diphosphate + L-glutamyl-tRNAGlu | - |
? | |
ATP + L-glutamate + tRNAGlu | preferred tRNA substrate | Thermosynechococcus vestitus | AMP + diphosphate + L-glutamyl-tRNAGlu | - |
? | |
additional information | the non-discriminating enzyme charges both tRNAGlu and tRNAGln with glutamate, anticodons of tRNAGlu, 34C/UUC36, and tRNAGln, 34C/UUG36, differ only in base 36, residue Gly366 is responsible for allowing both cytosine and the bulkier purine base G36 of tRNAGln to be tolerated, glutamate recognition structure, overview | Thermosynechococcus vestitus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structure comparison of non-discriminating GluRS versus the discriminating GluRS, EC 6.1.1.17, overview | Thermosynechococcus vestitus |
Synonyms | Comment | Organism |
---|---|---|
ND-GluRS | - |
Thermosynechococcus vestitus |
non-discriminating glutamyl-tRNA synthetase | - |
Thermosynechococcus vestitus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.036 | - |
tRNAGln | - |
Thermosynechococcus vestitus | |
0.1 | - |
tRNAGlu | - |
Thermosynechococcus vestitus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Thermosynechococcus vestitus |