7.6.2.8 purified recombinant complex of inhibitory nanobody Nb9 with enzyme BtuF complex, crystallization solution contains 100 mM Tris-HCl, pH 8.5, 400 mM MgCl2, and 33% w/v PEG4000, X-ray diffraction structrue determination and analysis at 2.7 A resolution 752215 7.6.2.8 purified recombinant His-tagged enzyme BtuM with natively bound cobalmin and anomalously bound cobalmin, sitting drop vapour diffusion method, mixing of 0.002 ml of protein in 50mM Tris-HCl, pH 7.5, or in 50 mM HEPES-NaOH pH 8.0, 100 mM NaCl, 0.005 mM cyano-Cbl and 0.35% detergent, with 0.002 ml of precipitant solution containing 25 mM Tris, pH 8.5, and 25-30% v/v PEG 400 or 50 mM Tris, pH 8.5, and 27-30% v/v PEG 400, or 75 mM Tris, pH 8.5, and 29-30% v/v PEG 400, at 8°C, 3-4 weeks, X-ray diffraction structure determination and analysis at 2.0-2.5 A resolution, structure modeling 751670 7.6.2.8 purified recombinant wild-type BtuCD-F, apo-BtuCD-F, and BtuCD-F mutant E159Q/N162C, X-ray diffraction structure determination and analysis 750063 7.6.2.8 vitamin B12-bound VcBtuF, protein in a solution with vitamin B12 in a 3:1 ratio, and 50 mM Tris-HCl, pH 7.0, and 300 mM NaCl, mixing of 0.003 ml of protein solution with 0.002 ml of precipitant solution containing 0.8 M ammonium sulfate, 0.1 M Tris, pH 8.0, and equilibration against a reservoir solution containing 0.5 ml of 1.6 M ammonium sulfate, 0.1 M HEPES, pH 7.0, 20°C, 7 days, hanging drop vapour diffusion method, X-ray diffraction structure determination and analysis at 1.67 A resolution 749978