    3.6.1.11 | purified recombinant detagged wild-type enzyme by hanging drop vapour diffusion, mixing of 0.002 ml of 10 mg/ml protein in 50 mM Tris, pH 8.0, 500 mM NaCl, and 5% glycerol, with 0.001 ml of reservoir solution containing 15.2% w/v PEG 3350, 2% w/v PEG 8000, 72 mM Tris, pH 8.0, 144 mM magnesium chloride hexahydrate, 20 mM HEPES/sodium hydroxide pH 7.5, 1.6% v/v glycerol, and 8% v/v DMSO, and equilibration against 0.2 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 3.3 A resolution. Purified recombinant detagged mutant truncated enzyme and truncated mutant E137A by sitting drop vapour diffusion, mixing of 0.001 ml of 8 mg/ml protein in 50 mM Tris, pH 8.0, 500 mM NaCl, and 5% glycerol, with 0.001 ml of reservoir solution containing 0.2 M magnesium chloride hexahydrate, 0.1 M Tris, pH 8.5 or pH 6.5, respectively, 25% w/v PEG 3350, and equilibration against 0.1 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 1.80 and 1.56 A resolution, respectively. Molecular replacement |
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