1.1.1.387 | purified recombinant wild-type and mutant enzymes in complex with NADP+/sulfate ion and with NADP+/L-tartrate (substrate analogue), sitting-drop vapor diffusion method, mixing of 0.001 ml of 13 mg/mL protein in 10 mM Tris-HCl, pH 8.0, 200 mM NaCl, and 5 mM 2-mercaptoethanol, with 0.001 ml of mother liquor containing 30% w/v PEG 2000 MME, 0.2 M ammonium sulfate, and 100 mM acetate buffer, pH 4.6, for complex crystals 0.5 mM NADP+ is added to the protein solution, soaking of the crystals in mother liquor containing 0.45 M L-(+)-tartaric acid for 10 min, X-ray diffraction structure determination and analysis at 1.18 and 1.57 A resolution, respectively. Based on the model of the selenomethionyl enzyme (PDB entry, 3W6U), the structure of the NADP+/ sulfate ion-bound and NADP+/ tartrate-bound enzymes is determined using the molecular replacement method, modeling |