EC Number   |
|---|
    6.3.2.3 | - |
| 6.3.2.3 | analysis of the dimeric enzyme, PDB ID 2HGS |
| 6.3.2.3 | at 2.0 A resolution |
| 6.3.2.3 | computational structure modeling of wild-type and mutant enzymes using crystal structure data at 2.1 A resolution, interactions of actie site residues Glu144, Asn146, Lys305, and Lys364, and ligands ADP2-, SO42-, and Mg2+ |
| 6.3.2.3 | crystallization by vapour-diffusion hanging-drop method of the free enzyme or the enzyme liganded to substrate gamma-glutamylcysteine and non-hydrolyzable ATP-substrate-analogue AMP-PNP, 17 mg/ml enzyme in 10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 1 mM DTT, plus equal volume of reservoir solution: for crystals of free enzyme with 1.97 M ammonium sulfate, 0.1 M Tris-HCl, pH 8.0, 2% PEG 400 at 22°C, or for the liganded enzyme with 3 mM AMP-PNP, 10 mM MgCl2, 3 mM gamma-glutamylcysteine against a well solution of 2.2 M ammonium sulfate, 0.1 M Tris-HCl, pH 8.0, 2% PEG 400, 5 mM TCEP, 40 mM MgCl2, at 22°C, X-ray diffraction structure determination and analysis at 2.3 A and 1.8 A, respectively |
| 6.3.2.3 | hanging drop vapour diffusion method |
| 6.3.2.3 | mutant enzymes replaced with Val at the basal position of the flexible loop (P227V, G240V, and P227V/G240V) are identical with wild-type enzyme in their crystal structures, except the loop region |
| 6.3.2.3 | purified recombinant enzyme, 20 mg/ml protein by hanging-drop method, from reservoir solution containing containing 10% w/v PEG 20K, 5% v/v tacsimate and 0.1 M HEPES, pH 7.5, 2-5 days, X-ray diffraction structure determination and analysis at 3.5 A resolution. Better crystals by vapor diffusion experiments to the without-oil microbatch method, method development, mixing of 40 mg/ml protein with an equal volume of solution containing 14% w/v PEG 20000, 0.2 M HEPES pH 7.5, 10% v/v tacsimate, X-ray diffraction structure determination and analysis at 2.34 A resolution, molecular replacement |
| 6.3.2.3 | structure modeling |
| 6.3.2.3 | structure of Escherichia coli B glutathione synthetase complexed with ADP, glutathione, and sulfate at 2.0 A resolution |
