    1.14.15.18 | purified recombinant wild-type Vdh and mutant Vdh-K1 in complex with substrates VD3 and 25(OH)VD3, 20 mg/ml wild-type protein in 20 mM Tris-HCl, pH 7.5, supplemented with 0-0.2% PMbetaCD for VD3 complex or 0-0.2% gammaCD for 25(OH)VD3 complex, 20 mg/ml mutant protein in 20 mM Tris-HCl, pH 7.5, supplemented with 20 mM NaCl, sitting-drop method, 20°C. Vdh-WT crystals are grown using reservoir solution containing 0.1 M BisTris, pH 7.5, 50 mM CaCl2, 40-120 mM NaCl or KCl, and 32-40% PEG 400 or PEG 550 monomethyl ether or 20% PEG 1000, Vdh-K1 is crystallized using reservoir solution containing 0.1 M calcium acetate and 10-14% PEG 3350, X-ray diffraction structure determination and analysis at 1.75-3.05 A resolution |
