EC Number   |
|---|
    2.3.2.2 | ammonium sulfate precipitation from 20 mM Tris-HCl, pH 8.0, 14.2 mg/ml protein, refrigerator, 1 week |
| 2.3.2.2 | ammonium sulfate precipitation, hanging-drop vapor diffusion method, crystal structure of gamma-glutamyltransferase at 1.95 A resolution and structure of the gamma-glutamyl-enzyme intermediate trapped by flash cooling the GGT crystal soaked in glutathione solution and the structure of GGT in complex with L-glutamate |
| 2.3.2.2 | ammonium sulfate precipitation, hanging-drop vapor diffusion method, crystal structure of the T391A mutant gamma-glutamyltranspeptidase that lacks autocatalytic processing ability refined at 2.55 A resolution |
| 2.3.2.2 | ammonium sulfate precipitation, refrigerator, 2 weeks |
| 2.3.2.2 | in complex with azaserine and acivicin, at 1.65 A resolution. Both inhibitors bind to the substrate-binding pocketand form a covalent bond with the Ogamma atom of residue T391. The two amido nitrogen atoms of Gly483 and Gly484, which form the oxyanion hole, interact with the inhibitors directly or via a water molecule. In the azaserine complex the carbon atom that forms a covalent bond with Thr391 is sp3-hybridized |
| 2.3.2.2 | in complex with L-glutamate, hanging drop vapor diffusion method, using poly(ethylene glycol) 4000, 100 mM MES buffer (pH 7.0), 600 mM NaCl, and 5% (v/v) Jeffamine M-600 |
| 2.3.2.2 | purified recombinant deglycosylated hGGT1 mutant V272A with bound inhibitor DON, mixing of 0.002 ml of 4.3 mg/ml protein in 50 mM HEPES, pH 8.0, 0.5 mM EDTA, 0.02% sodium azide, and 2 mM DON, with 0.0017 ml of H2O, and 0.002 ml of reservoir solution containing 20-25% PEG 3350, 0.1 M Na-cacodylate, pH 6.0, and 0.1 M ammonium chloride, microseeding with apo-hGGT1 crystals, room temperature, 1-2 days, X-ray diffraction structure determination and analysis at 2.20 A resolution, modeling using the apo-form crystals of hGGT1 (PDB ID 4Z9O) as template |
| 2.3.2.2 | purified recombinant wild-type PnGGT enzyme, sitting drop vapor diffusion method, mixing of 0.001 ml of 3-5 mg/ml protein in 0.1 M HEPES, pH 7.0, with 0.001 ml reservoir solution containing 16% PEG 8000, 15% PEG 400, 0.1 M HEPES, pH 7.0, 50 mM glycylglycine, and equilibration against 0.5 ml of reservoir solution, 20°C, 1 week, method optimization, X-ray diffraction structure determination and analysis at 1.57-1.70 A resolution, molecular replacement method using the structure of Escherichia coli GGT (EcGGT, PDB ID 2DG5) as the template, modeling |
