   3.4.22.1 | purified wild-type enzyme and enzyme mutants in complex with inhibitor chagasin, hanging drop vapor diffusion method at 18°C, for crystal form I: mixing of 0.001 ml of 5 mg/ml chagasin solution with 0.002 ml of protein solution containing 3 mg/ml cathepsin B, and 0.002 ml of precipitant solution containing 0.2 M NH4H2PO4, 20% PEG 3350, pH 4.6, 2 weeks, for crystal form II: mixing of 0.001 mL of 5 mg/ml chagasin solution with 0.0015 ml of protein solution containing 3 mg/ml cathepsin B, and 0.001 ml of precipitant solution containing 0.14 M NH4F, 14% PEG 3350, pH 6.2, 1 week, X-ray diffraction structure determination and analysis at 1.8-2.7 A resolution |
