NINDS INSTITUTIONAL CENTER CORE GRANTS - Neuroproteomics P30

这项提议的目的是建立一个华盛顿大学研究核心设施，为一大批对绘制神经系统中蛋白质-蛋白质相互作用图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，对其结合伙伴的明确分析将有助于研究人员了解这些基因的变化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NINDS的任务至关重要。三个研究核心和一个行政核心将为研究方案提供支持，并将促进调查人员之间的互动和合作。核心A，分子病毒学核心，将为标记蛋白质和绘制蛋白质-蛋白质相互作用图谱以及使用各种生化方法验证结合相互作用所需的所有表达质粒提供一个储存库。这个核心还将为每个表达构建物生成和滴定慢病毒，为在传统上难以转化的细胞中表达标记蛋白提供了宝贵的试剂。核心B，即质谱学核心，将提供对使用与核心A中产生的各种标记蛋白质相关的下拉技术提纯的蛋白质复合体的基于质谱学的鉴定。核心C，生物信息学核心，将提供对各种下拉分析中识别的蛋白质的蛋白质序列数据、分子相互作用、途径、生物结构、遗传图谱、同源性信息和功能注释的全面分析。研究核心将由一个行政核心提供支助，该行政核心将监督每个研究核心的运作，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，并编写提交给供资机构的报告。

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.