Structure Determination by NMR

NMR 结构测定

• 批准号: 6495130
• 负责人: DAVID E WEMMER
• 金额: $ 109.44万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6495130
• 关键词: Escherichia coli; Mycoplasma; bacterial proteins; chemical information system; high throughput technology; mycoplasmal pneumonia; nuclear magnetic resonance spectroscopy; protein folding; protein structure; protein structure function; thermophilic organism

The overall project will determine the structure of proteins from Mycoplasma genitalium and Mycoplasma pneumoniae. These structures will enhance the database of protein folds, and will be interpreted to suggest possible functions for the proteins. This component of the overall project will apply solution NMR methods to determine structures of small proteins from these organisms using multinuclear multidimensional methods. Sets of experiments will be designed which are optimized for assignment of resonances from proteins with specific linewidths. Methods for automated analysis of data (determination of backbone and sidechain assignments, and assignment of NOE crosspeaks) will be implemented, and extended to optimize throughput. Proteins homologous to M.g. and M.p. but from thermophilic organisms will be evaluated fro NMR work. Data collection at higher temperature is improved due to decreased linewidths, enhancing data quality and hence the ability to automate analysis. The interpretation of structures determined in terms of fold classification and function will be done in collaboration with other members of the project. NMR will also be applied to evaluation of proteins for crystallography, attempting to identify flexible regions, and comains which may hinder crystallography, attempting to identify flexible regions, and domains which may hinder crystallization.

整个项目将确定生殖支原体和肺炎支原体蛋白质的结构。这些结构将增强蛋白质折叠的数据库，并将被解释为蛋白质可能的功能。整个项目的这一部分将应用溶液核磁共振方法，使用多核多维方法确定这些生物体中小蛋白的结构。将设计一系列实验，优化分配具有特定线宽的蛋白质的共振。将实施和推广数据自动分析方法(确定主干和侧链分配，以及NOE串音分配)，以优化吞吐量。与M.G.同源的蛋白质。和M.P.但来自嗜热生物的物质将通过核磁共振工作进行评估。由于减少了线宽，提高了数据质量，从而提高了自动化分析的能力，从而改善了在较高温度下的数据收集。将与该项目的其他成员合作，对根据褶皱分类和功能确定的结构进行解释。核磁共振还将应用于蛋白质结晶学的评估，试图确定可能阻碍结晶学的柔性区域和常见区域，试图确定可能阻碍结晶的柔性区域和结构域。