EXOMER COMPLEX

外显子复合体

• 批准号: 8363377
• 负责人: J Christopher Fromme
• 金额: $ 0.29万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363377
• 关键词: Adaptor Signaling Protein; Binding; Biochemical; Biological Assay; Capsid Proteins; Carrier Proteins; Cell Cycle; Cell membrane; Clathrin Adaptors; Complex; Crystallography; Data Set; Devices; Funding; Grant; Home environment; L-Selenomethionine; Light; Membrane; Modeling; Monitor; Names; National Center for Research Resources; Play; Principal Investigator; Process; Protein Binding; Proteins; Research; Research Infrastructure; Resort; Resources; Roentgen Rays; Role; Shapes; Source; Stress; Synchrotrons; System; United States National Institutes of Health; Vesicle; Yeasts; beamline; cost; interest; response; trafficking; trans-Golgi Network

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. Transport of proteins between membrane-bound compartments is a tightly regulated process involving vesicular membrane carriers. A classical membrane vesicle is created by membrane deformation achieved in part by cytosolic "coat" proteins. Coat proteins bind and shape membranes, and they also play an important role in selecting cargo incorporated into transport carriers. Recent studies have established the existence of a new membrane coat complex responsible for trafficking certain cargo proteins from the trans-Golgi network (TGN) to the plasma membrane in yeast. This coat, named "Exomer", is interesting as an alternative to the better-known TGN coat systems involving Clathrin and Adaptor Proteins. Exomer is also quite interesting as a model for regulated transport. Under steady-state conditions, cargo trafficked by exomer is localized in a stable internal compartment, but in response to stress or progression through the cell-cycle cargo is rapidly mobilized to the plasma membrane in a polarized manner. The proper localization of two Exomer cargo proteins in particular (Chs3 and Fus1) is easily monitored by simple assays, making this an excellent system for in-depth mechanistic studies. We are pursuing structural and biochemical studies of Exomer in order to understand how it functions. We currently have diffracting crystals of a fully functional exomer complex, having collected a native 2.8A dataset on the CHESS A1 beamlime. We are unable to grow crystals of the SeMet-substituted protein, so we are now resorting to heavy-atom soaks for MIR/MIRAS/SIRAS. We are unable to screen for derivatives on a home source because our crystals do not diffract strongly enough using home-source X-rays. An insertion device beamline is ideal, as the best we can do is 2.8A at CHESS A1. We also would like to take advantage of tunable wavelength for attempts at MIRAS/SIRAS.

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。 蛋白质在膜结合的隔室之间的转运是一个涉及囊膜载体的严格调控的过程。经典的膜泡是由膜变形形成的，部分是由细胞质的“外壳”蛋白实现的。外壳蛋白结合和塑造膜，它们在选择纳入运输载体的货物方面也发挥着重要作用。最近的研究证实，酵母中存在一种新的膜包膜复合体，负责将某些货物蛋白从反式高尔基网络(TGN)运输到质膜。这种被命名为“Exmer”的外衣很有趣，因为它是一种更知名的TGN外衣系统的替代品，它涉及笼状蛋白和接头蛋白。Exmer作为受管制运输的模型也相当有趣。在稳态条件下，外显体运输的货物被定位在一个稳定的内部隔室，但为了响应压力或在细胞周期中的进展，货物以极化的方式迅速动员到质膜上。两个外聚体货物蛋白(Chs3和FUS1)的正确定位很容易通过简单的分析进行监测，这使其成为深入机制研究的优秀系统。我们正在进行Exmer的结构和生化研究，以了解它的功能。我们目前拥有一个功能齐全的外聚体复合体的衍射晶体，已经收集了国际象棋A1光束石灰上的本地2.8A数据集。我们无法生长SeMet取代的蛋白质的晶体，所以我们现在求助于重原子浸泡来获得MIR/MIRAS/SIRAS。我们无法在原生源上筛选衍生品，因为我们的晶体使用原生源X射线的衍射力不够强。插入装置光束线是理想的，因为我们在国际象棋A1中最多能做到2.8A。我们还希望利用可调波长来尝试MIRAS/SIRAS。