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Crystallization and preliminary X-ray analysis of cholesterol oxidase from Brevibacterium sterolicum containing
1Biochemistry Department, Montreal Joint Centre for Structural Biology, McIntyre Medical Sciences Building, McGill University, 3655 Drummond Street, Montréal, Québec, H3G 1Y6, Canada.
Abstract:
Single crystals of cholesterol oxidase from Brevibacterium sterolicum containing a covalently bound form of the FAD cofactor have been obtained. The crystals are grown by vapor diffusion using the hanging drop technique from 12% polyethylene glycol, Mr 8000, and 75 mM MnSO4 as the precipitant at pH 5.2. In order to obtain large diffraction quality crystals, nucleation must occur at 22 degrees C with subsequent growth at 17 degrees C. The crystals belong to the monoclinic space group P21 with cell dimensions a = 78. 5 A, b = 126.7 A, c = 82.4 A and beta = 108.9 degrees with two protein molecules per asymmetric unit. Diffraction of these crystals has been observed to at least 2.2 A resolution and they are suitable for an X-ray structure analysis.