  1.13.11.78 | purified enzyme in complex with substrate (R)-2-amino-1-hydroxyethylphosphonate or with L-tartrate, derived from the crystallization buffer, hanging drop vapour diffusion method, mixing 0.005 ml of 16 mg/ml selenomethionine-labeled protein of enzyme in complex with L-tartrate in a solution containing 5% n-octyl-beta-D-glucoside with 0.005 ml of reservoir solution containing 2.4 M ammonium sulfate and 0.15 M potassium sodium L-tartrate, 5-7 days at room temperature, the substrate-complexed enzyme is crystallized by sitting drop vapour diffusion method by mixing of 0.001 ml of protein solution with 0.001 ml of reservoir solution containing 0.1 M Bis-Tris, pH 6.5 and 20% w/v PEG 5000 monomethyl ether, X-ray diffraction structure determination and analysis at 2.1 and 1.7 A resolution, respectively, molecular replacement and modeling |
