MOLECULAR ENVELOPE RECONSTRUCTIONS OF THE EXOMER CARGO ADAPTOR COMPLEX

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 小的GTPase Arf1如何招募蛋白质转运货物适配器到膜上的基础在很大程度上仍不清楚。虽然Arf1的结构已经知道多年，但我们对Arf蛋白如何相对于膜或膜结合的货物蛋白定位货物适配器或囊泡外壳知之甚少。Exmer是一种受时间调控的Arf1依赖的寡聚体货物适配器。有趣的是，其依赖于Arf1的膜结合需要外聚体的齐聚。我们计划表征外聚体复合体的结构以及它如何与Arf1结合，以阐明Arf1结合在货物适配子膜结合中的作用。 在CHESS上进行的SAXS实验将用于确定外聚体的低聚状态，并用于定位复合体中的亚基。此外，对Arf1-外聚体复合体的SAXS分析将表明Arf1结合在外聚体低聚物上的位置。

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. The basis for how the small GTPase Arf1 recruits protein trafficking cargo adaptors to membranes remains largely unknown. Although the structure of Arf1 has been known for years, we know very little about how Arf proteins orient cargo adaptors or vesicle coats with respect to membranes or membrane-bound cargo proteins. Exomer is a temporally regulated Arf1-dependent oligomeric cargo adaptor. Interestingly, oligomerization of exomer is required for its Arf1-dependent membrane binding. We plan to characterize the architecture of the exomer complex and how it binds to Arf1, in order to elucidate the role of Arf1-binding in cargo adapter membrane association. SAXS experiments performed at CHESS will be used for determining the oligomeric state of exomer, and for positioning the subunits within the complex. Furthermore, SAXS analysis of an Arf1-exomer complex will indicate the position of Arf1-binding on the exomer oligomer.