来自含有共价结合黄素腺嘌呤二核苷酸（FAD）的短杆菌属胆固醇氧化酶的结晶及初步X射线分析。

Crystallization and preliminary X-ray analysis of cholesterol oxidase from Brevibacterium sterolicum containing covalently bound FAD.

作者信息

Croteau N, Vrielink A

机构信息

Biochemistry Department, Montreal Joint Centre for Structural Biology, McIntyre Medical Sciences Building, McGill University, 3655 Drummond Street, Montréal, Québec, H3G 1Y6, Canada.

出版信息

J Struct Biol. 1996 Mar-Apr;116(2):317-9. doi: 10.1006/jsbi.1996.0047.

DOI:10.1006/jsbi.1996.0047

PMID:8812988

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.

摘要

已获得来自短杆菌属的胆固醇氧化酶的单晶，其中含有共价结合形式的黄素腺嘌呤二核苷酸（FAD）辅因子。通过悬滴法，以12%聚乙二醇（分子量8000）和75 mM硫酸锰作为沉淀剂，在pH 5.2条件下通过气相扩散法生长晶体。为了获得高质量的衍射晶体，成核必须在22℃下发生，随后在17℃下生长。这些晶体属于单斜空间群P21，晶胞参数为a = 78.5 Å，b = 126.7 Å，c = 82.4 Å，β = 108.9°，每个不对称单元中有两个蛋白质分子。已观察到这些晶体的衍射分辨率至少为2.2 Å，适合进行X射线结构分析。