Cloned (Comment) | Organism |
---|---|
expression of Glu-Q-RS in Escherichia coli strain BL21(DE3) | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
Glu-QRS complexed to Glu, sitting drop vapour diffusion method, mixing of 0.002 ml of protein solution, containing 9.7 mg/ml protein in 20 mM Na-HEPES buffer, pH 7.2, and 50 mM NaCl, with 0.002 ml of reservoir solution containing 0.1 M Mg-acetate and Na-cacodylate buffer, pH 5.5, 0.2 M KCl, 10% polyethylene glycol 8000, and 2 mM L-Glu, a few days, X-ray diffraction structure determination and analysis at 1.5 A resolution | Escherichia coli |
General Stability | Organism |
---|---|
presence of 10% glycerol during the purification prevents the precipitation of Glu-Q-RS | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics and binding constants | 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 + tRNAAsp | Escherichia coli | the L-glutamyl-queuosine tRNAAsp synthetase, Glu-Q-RS from Escherichia coli is a paralogue of the catalytic core of glutamyl-tRNA synthetase, GluRS, that catalyzes glutamylation of queuosine in the wobble position of tRNAAsp | AMP + diphosphate + L-glutamyl-tRNAAsp | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant Glu-Q-RS from Escherichia coli strain BL21(DE3) by anion exchange and hydroxyapatite chromatography, followed by dialysis, presence of 10% glycerol during the purification prevents the precipitation of Glu-Q-RS | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate + tRNAAsp | the L-glutamyl-queuosine tRNAAsp synthetase, Glu-Q-RS from Escherichia coli is a paralogue of the catalytic core of glutamyl-tRNA synthetase, GluRS, that catalyzes glutamylation of queuosine in the wobble position of tRNAAsp | Escherichia coli | AMP + diphosphate + L-glutamyl-tRNAAsp | - |
? | |
ATP + L-glutamate + tRNAAsp | the L-glutamyl-queuosine tRNAAsp synthetase, Glu-Q-RS from Escherichia coli is a paralogue of the catalytic core of glutamyl-tRNA synthetase, GluRS, that catalyzes glutamylation of queuosine in the wobble position of tRNAAsp. Activation of Glu to form Glu-AMP, the intermediate of tRNA aminoacylation, in the absence of tRNA. Glu-Q-RS transfers the activated Glu to Q34 located in the anticodon loop of the noncognate tRNAAsp. A C in position 38 is crucial for glutamylation of Q34 | Escherichia coli | AMP + diphosphate + L-glutamyl-tRNAAsp | - |
? | |
additional information | Glu-Q-RS binds the noncognate amino acids L-Gln and D-Glu fourfold and sixfold, respectively, weaker than L-Glu. Despite important structural similarities, Glu-Q-RS and GluRS diverge strongly by their functional properties, selection of the cognate amino acid and by the mechanism of its activation, overview. Structural basis of the reaction mechanism, overview | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | three-dimensional structure of Glu-QRS complexed to Glu, the enzyme lacks the anticodon-binding domain | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
Glu-Q-RS | - |
Escherichia coli |
glutamyl-queuosine tRNAAsp synthetase | - |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | binding site pocket structure, overview | Escherichia coli |