CRYSTALLIZATION AND X RAY CRYSTALLOGRAPHY

蛋白质结晶和X射线核心的具体目标 为结晶学提供必要的设施和技术 包括人员在内的资源，用于确定 X射线测定PGHS同工酶、细菌细胞色素C氧化酶及其突变体 衍射技术。核心将提供以下内容： 1.用于结晶的纯化蛋白质样品的制备这将是 包括协助或直接参与洗涤剂交换， 在透析中，在制备特定的蛋白质-配体复合体中 用于结晶。 2-结晶筛的设计、开发和安装 该计划项目中的特定膜蛋白。这将包括 适合结晶的新型洗涤剂体系的筛选 条件。 3.晶体的表征(如：十二烷基硫酸钠-PAGE、晶体密度等) 包括初步的X射线表征，然后是所需的 生长X射线质量的结晶方案的优化 水晶。 5.X射线核心人员将设计和执行X射线数据收集 和数据处理，尽管在适当的时候也将有助于其他 项目成员在数据收集和处理方面。 6.X光核心人员将与计划成员合作 晶体结构分析项目以及提供任何所需的 为确定结构提供的技术援助和专业知识 计划项目的个别成员。

The specific aims of the Core for Protein Crystallization and X-ray Crystallography are to provide the necessary facilities and technical resources, including personnel, for the structure determination of the PGHS isozymes, bacterial cytochrome c oxidase and mutants thereof by X-ray diffraction techniques. The core will provide the following: 1. Preparation of purified protein samples for crystallization. This will include either assistance in or direct involvement in detergent exchange, in dialysis, and in the preparation of particular protein-ligand complexes for crystallization. 2- Design, development and setup of crystallization screens for the specific membrane proteins in the program project. This will include the screening of novel detergent systems for suitable crystallization conditions. 3. Characterization of crystals (e.g., SDS-PAGE, crystal density, etc.) including preliminary X-ray characterization, followed by any needed optimization for the crystallization protocols to grow X-ray quality crystals. 5. The X-ray Core personnel will design and execute X-ray data collection and data processing although, when appropriate, will also assist other members of the program project in data collection and processing. 6. The X-ray Core personnel will collaborate with members of the program project in the crystal structure analysis as well as provide any needed technical assistance and expertise for structure determination to individual members of the program project.