3.1.3.75 | purified recombinant enzyme, sitting drop vapour diffusion method, 250 nl of 10 mg/ml protein in 200 mM NaCl, 10 mM Tris-HCl, pH 8.0, are mixed with 250 nl well solution and are equilibrated against 0.5 ml well solution containing 0.1 M Bis-Tris, pH 5.5, 0.2 M MgCl2, 25% PEG 3350, producing clustered plate-like microcrystals, or by microbatch method mixing 0.001 ml of protein solution containing 10 mg/ml in 200 mM NaCl, 20 mM Bis-Tris, pH 5.5, with 0.002 ml precipitant solution, cryoprotection by a quick soak in 0.1 M Bis-Tris pH 5.5, 0.2 M MgCl2, 25% PEG 3350, 15% glycerol, X-ray diffraction structure determination and analysis at 2.7 A resolution |
713654 |