   1.1.1.370 | purified His-tagged wild-type and selenomethionine-labeled enzyme in apoform and in a ternary complex with NAD+ and L-glucono-1,5-lactone, and in complex with myo-inositol and scyllo-inosose, hanging-drop vapor-diffusion method, mixing of 0.002 ml of protein solution with 0.02 ml of reservoir solution containing 0.1 M sodium acetate pH 4.8, and 12% PEG 3350, 20°C, crystals are soaked for one minute with 25 mM of NAD+ and 100 mM substrate in the reservoir solution, X-ray diffraction structure deteramintion and analysis at 1.75-2.3 A resolution, the structures complexed with myo-inositol/NAD+ and scyllo-inosose/NAD+ are solved at 2.3 A and 2.0 A resolution, respectively. Molecular replacement using the Se-Met-structure as the search model, modeling. The crystal is soaked with L-glucose and the resulting structure is determined |
