T. brucei Mitochondrial Proteome

描述（由申请人提供）：本项目将表征布氏锥虫的线粒体（mt）蛋白质组，以及生命周期阶段之间的蛋白质差异。它将阐明潜在的所有mt蛋白质的功能，并构建结构和功能蛋白质相互作用网络。这将扩展对mt生理学的理解，并确定潜在的药物靶点。该项目将：1。使用高通量串联质谱（MS/MS）、SBEAMs数据库/数据管理系统和一套分析软件，尽可能多地鉴定前环形式（PF）mt及其组分中的蛋白质。和Web接口。分馏将降低样品的复杂性，增强肽检测，并确定亚细胞器的位置和多蛋白复合物。2.通过鉴定纯化的ml复合物的蛋白质组成来确定线粒体蛋白质之间的关联。许多蛋白质的一般功能将建议从复合物中的共定位。3.构建线粒体蛋白质相互作用组。目标1和2的数据集将与使用SBEAMS模块的外部来源的大量数据的计算机分析相结合，从中将生成物理和功能相互作用网络，使用CYTOSCAPE进行可视化，并通过网络传播。这将生成T的配置文件。布氏结核病生理学，并确定其途径和过程中的关键步骤。4.比较血流和前循环形式的线粒体蛋白质组。将在血流（BF）和PF之间比较mt复合物的存在和组成。这些阶段之间总mt蛋白的差异将使用aim 1中的程序进行调查。这将为BF生理学提供见解，并有助于预测必需基因。5.检查基因失活和药物治疗的后果。将分析具有预测关键功能的基因失活的效果。这将评估复杂的共定位，相互作用图的有效性，并评估关键蛋白质的一般功能以及药物敏感性。总的来说，该项目将扩展TriTryp基因组注释，阐明mt功能组织及其在生命周期阶段之间的差异，并帮助鉴定mt药物靶标和影响药物敏感性的因素。

DESCRIPTION (provided by the applicant): This project will characterize the mitochondrial (mt) proteome of Trypanosoma brucei, and protein differences between life cycle stages. It will elucidate the functions of potentially all mt proteins and construct structural and functional protein interaction networks. This will extend understanding of mt physiology, and identify potential drug targets. The project will: 1. Identify as many proteins as possible in procyclic form (PF) mt and fractions thereof using high-throughput tandem mass spectroscopy (MS/MS), the SBEAMs data base/data management system, a suite of analytical software. and web interfaces. Fractionation will reduce sample complexity, enhance peptide detection, and identify sub-organellar locations and multi-protein complexes. 2. Determine associations among mitochondrial proteins by identifying the protein composition of purified ml complexes. The general functions of numerous proteins will be suggested from co-localization in complexes. 3. Construct the mitochondrial protein interactome. The datasets from aims 1 and 2 will be integrated with in silico analyses of extensive data from external sources using SBEAMS modules from which physical and functional interaction networks that will be generated, visualized using CYTOSCAPE, and disseminated via the web. This will generate a profile of T. brucei mt physiology and identify key steps in its pathways and processes. 4. Compare bloodstream and procyclic form mitochondrial proteomes. The presence of and composition of mt complexes will be compared between bloodstream (BF) and PFs. Differences in total mt proteins between these stages will be surveyed using procedures from aim 1. This will provide insights into BF physiology and be useful for predicting essential genes. 5.Examine the consequences of gene inactivation and drug treatment. The effect of inactivating genes with predicted critical functions will be analyzed. This will assess the validity of complex co-localizations, interaction maps, and assess the general functions of key proteins as well as drug susceptibility. Overall, this project will extend the TriTryp genome annotation, elucidate mt functional organization and its differences between life cycle stages, and aid the identification of mt drug targets and factors affecting drug susceptibility.