NINDS INSTITUTIONAL CENTER CORE GRANTS - Neuroproteomics P30

描述（由申请人提供）：本提案的目的是建立华盛顿大学研究核心设施，为大量对神经系统中蛋白质-蛋白质相互作用图谱感兴趣的科学家提供必要的蛋白质组学和生物信息学基础设施支持。这些研究人员中的许多人进行涉及疾病基因的研究，并对其结合伙伴进行明确的分析，将有助于研究人员了解这些基因的改变如何在神经系统疾病和损伤过程中调节神经元和神经胶质的结构和功能。这项研究显然对国家海洋学研究所的任务至关重要。三个研究核心和一个行政核心将为研究项目提供支持，并将促进研究人员之间的互动和合作。核心A，分子病毒学核心，将提供所有表达质粒的存储库，用于标记蛋白质和绘制蛋白质-蛋白质相互作用，并使用各种生化方法验证结合相互作用。该核心还将为每个表达构建体生成和滴度慢病毒，为在传统上难以转染的细胞中表达标记蛋白提供有价值的试剂。Core B是质谱核心，将提供基于质谱的蛋白质复合物鉴定，这些蛋白质复合物使用与Core a中生成的各种标记蛋白质相关的pull - down技术纯化。Core C是生物信息学核心，将提供蛋白质序列数据、分子相互作用、途径、生物结构、遗传图谱、同源信息和各种pull - down测定中鉴定的蛋白质的功能注释的综合分析。各研究中心将由行政中心提供支援，行政中心将监督各研究中心的运作、组织用户会议、指导委员会会议、组织年度研究专题讨论会、提供文书、财政和人事支助，并向资助机构编写报告。

DESCRIPTION (provided by applicant): The purpose of this proposal is to establish a University of Washington Research Core facility that provides essential proteomics and bioinformatics infrastructure support for a large contingent of scientists interested in mapping protein-protein interactions in the nervous system. Many of these investigators conduct research involving disease genes and a defined analysis of their binding partners will help investigators to understand how alterations in these genes modulate neuronal and glial structure and function during the course of nervous system disease and injury. This research is clearly essential to the mission of the NINDS. Three research cores and one administrative core will provide support for the research programs and will stimulate interactions and collaborations among investigators. Core A, the molecular virology core, will provide a repository for all of the expression plasmids needed to tag proteins and map protein-protein interactions and validate binding interactions using a variety of biochemical methods. This core will also generate and titer lentivirus for each expression construct providing a valuable reagent for expressing tagged proteins in cells that are traditionally difficult to transfect. Core B, the mass spectrometry core, will provide mass spectrometry-based identification of protein complexes purified using the pull down techniques associated with the various tagged proteins generated in Core A. Core C, the bioinformatics core, will provide comprehensive analysis of protein sequence data, molecular interactions, pathways, biological structures, genetic maps, homology information, and functional annotations of proteins identified in the various pull down assays. The research cores will be supported by an administrative core that will oversee the operation of each research core, organize user meetings, steering committee meetings, organize an annual research symposium, provide clerical, fiscal and personnel support, and prepare reports to the funding agency.