CHARACTERIZATION OF MEMBRANE PROTEIN COMPLEXES

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该项目的重点是发展和使用透射电子显微镜在 表征膜蛋白-蛋白复合物。 我们将开发一种自动化检测方法， 根据膜蛋白复合物聚集状态对其进行成像、选择和分类， 尺寸、构象状态和结构域排列。一个目标是开发一个更 系统了解去污剂和脂质对稳定性的影响， 膜蛋白及其复合物的构象，以帮助优化 结晶的努力。 一个更雄心勃勃的目标是绘制出单个蛋白质结构域， 为了更好地理解蛋白质之间的相互作用， 构象上或结构上不均匀的。 这种低分辨率信息可以是 与单个亚基或亚复合体的高分辨率模型相结合， X射线和核磁共振研究，以发展更详细的了解蛋白质- 复合物中的蛋白质相互作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This project focuses on the development and use of transmission electron microscopy in characterizing membrane protein-protein complexes. We will develop an automated assay to image, select, and classify membrane protein complexes in terms of their aggregation state, size, conformational state and domain arrangement. One goal is to develop a more systematic understanding of the effect of detergents and lipids on the stability and conformation of membrane proteins and their complexes in order to help optimize crystallization efforts. A more ambitious goal is to map out individual protein domains in order to better understand the interactions between proteins in a complex that might be conformationally or structurally heterogeneous. This low resolution information can be combined with high-resolution models of individual subunits or subcomplexes, derived from X-ray and NMR studies, in order to develop a more detailed understanding of the protein- protein interactions within complexes.