   3.4.22.37 | purified enzyme gingipain R2 in complex with fast acting, irreversible inhibitor H-D-Phe-Phe-Arg-chloromethylketone, activated by incubation for 30 min in 0.1 M HEPES, pH 8.0, 10 mM L-cysteine, 2.5 mM CaCl2, at 37°C, alkylation of the active site cysteine by addition of inhibitor, crystallization by sitting-drop vapour diffusion method at 20°C, different protein-inhibitor complex solution systems using PEG 8000 as precipitant for 10 mg/ml protein, X-ray diffraction structure determination at 2.9 A resolution and analysis |
| 3.4.22.37 | purified RgpB free or in complex with fast acting, irreversible inhibitor D-Phe-Phe-Arg-chloromethylketone, activated by incubation for 30 min in 0.1 M HEPES, pH 8.0, 10 mM L-cysteine, 2.5 mM CaCl2, at 37°C, alkylation of the active site cysteine by addition of inhibitor, crystallization of dialysed sample by vapour diffusion method, 8 mg/ml protein in 3 mM MOPS, pH 7.2 0.02% NaN3, plus equal volume of reservoir solution: 3.4 M 1,6-hexandiol, 0.2 M MgCl2, 0.1 M Tris-HCl, pH 8.5, at 21°C, several weeks, X-ray diffraction structure determination at 1.5-2.16 A resolution and analysis, structure modeling |
