   1.13.12.19 | purified recombinant enzyme in three structures: of enzyme in complex with manganese and 2-oxoglutarate, of enzyme in complex with with manganese and bis-Tris-propane buffer, and enzyme in complex with iron, L-Arg, and N-oxalylglycine, hanging drop vapour diffusion method, mixing of 0.003 ml of 12-20 mg/ml protein solution with 0.003 ml of reservoir solution containing 17.5-25% w/v PEG 3350 and PEG 6000, 0.1 M bis-Tris-propane, pH 7.0, 0.04 M sodium/potassium phosphate, 3-5 mM MnCl2, and with or without 10 mM 2-oxoglutarate, and equilibration against 0.5 ml reservoir solution, at 20°, microseeding, method optimization, X-ray diffraction structure determination and analysis at 1.08-1.55 A resolution, single-wavelength anomalous diffraction data obtained from selenomethionine-derivatized PsEFE:Mn:bis-Tris-propane crystals, structure modeling |
