3.1.4.45 | purified recombinant detagged wild-type and selenomethionine-labeled enzyme, sitting nanodroplet vapor diffusion method, mixing of 200 nl of 21 mg/ml protein in 20 mM HEPES, pH 8.0, 200 mM NaCl, 40 mM imidazole, and 1 mM TCEP, with 200 nl of reservoir solution containing 0.2 M Li2SO4, 30% PEG 4000, and 0.1 M Tris, pH 8.5, and equilibration against 0.05 ml of reservoir solution, 4°C, 10% v/v ethylene glycol as a cryoprotectant, X-ray diffraction structure determination and analysis at 1.80 A resolution, modeling, the crystal structure of the BACOVA_00430 protein from Bacteroides ovatus is used to model the luminal portion of the human enzyme N-acetylglucosamine-1-phosphodiester alpha-N-acetylglucosaminidase |
730014 |