CRYSTALLIZATION AND X RAY CRYSTALLOGRAPHY

结晶和 X 射线晶体学

• 批准号: 6316677
• 负责人: R Michael Garavito
• 金额: $ 10.47万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6316677
• 关键词: SDS polyacrylamide gel electrophoresis; X ray crystallography; bioimaging /biomedical imaging; biomedical facility; cytochrome c; cytochrome c peroxidase; data collection; isozymes; membrane proteins; prostaglandin endoperoxide synthase; protein purification; structural biology; technology /technique development

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.

蛋白质结晶和X射线核心的具体目标 晶体学是提供必要的设施和技术 包括人员在内的资源，以确定 PGHS同工酶、细菌细胞色素c氧化酶及其突变体的X射线分析 衍射技术核心将提供以下内容： 1.用于结晶的纯化蛋白样品的制备。这将 包括协助或直接参与洗涤剂交换， 在透析和制备特定蛋白质-配体复合物中 用于结晶。 2-设计、开发和安装结晶筛 特定的膜蛋白在程序项目。这将包括 筛选适合结晶的新型洗涤剂体系 条件 3.晶体的表征（例如，SDS-PAGE、晶体密度等） 包括初步的X射线表征，然后是任何需要的 优化结晶方案以提高X射线质量 晶体 5. X射线核心人员将设计并执行X射线数据收集 数据处理，虽然，在适当的时候，也将有助于其他 对项目成员进行数据收集和处理。 6. X射线核心人员将与项目成员合作 项目中的晶体结构分析以及提供任何需要的 技术援助和专门知识， 项目的个人成员。