T. brucei Mitochondrial Proteome

描述(由申请人提供)：这个项目将描述布氏锥虫线粒体(MT)蛋白质组，以及不同生命周期阶段的蛋白质差异。它将阐明潜在的所有mt蛋白的功能，并构建结构和功能蛋白相互作用网络。这将扩大对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学(MS/MS)、SBEAMS数据库/数据管理系统、一套分析软件来识别尽可能多的原环(PF)mt及其部分蛋白质。和Web界面。分级分离将降低样品的复杂性，增强多肽检测，并识别亚细胞器位置和多蛋白质复合体。2.通过鉴定纯化的ML复合体的蛋白质组成，确定线粒体蛋白质之间的联系。许多蛋白质的一般功能将从复合体中的共定位中得到推测。3.构建线粒体蛋白质互作组。来自AIMS 1和2的数据集将与使用SBEAMS模块对来自外部来源的大量数据进行电子分析相结合，这些模块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。这将生成布氏支原体的生理学概况，并确定其途径和过程中的关键步骤。4.比较血流和原环状线粒体蛋白质组。我们将比较血流(BF)和PFS中MT复合体的存在和组成。这些阶段之间的总mt蛋白的差异将使用目标1中的程序进行调查。这将提供对BF生理学的见解，并有助于预测必要的基因。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。总体而言，该项目将扩展TriTryp基因组注释，阐明mt的功能组织及其在生命周期阶段的差异，并帮助识别mt药物靶点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.