Mitochondrial Response to Oxidative Stress

线粒体对氧化应激的反应

• 批准号: 6771879
• 负责人: DANIEL F. BOGENHAGEN
• 金额: $ 37.63万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-10 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6771879
• 关键词: DNA repair; DNA replication; SDS polyacrylamide gel electrophoresis; cell differentiation; chromatography; electron transport; environmental exposure; epitope mapping; free radical oxygen; green fluorescent proteins; lipid peroxides; mass spectrometry; menadione; methylphenyltetrahydropyridine; mitochondrial DNA; mitochondrial membrane; oxidative stress; plant insecticide; protein localization; protein structure function; tissue /cell culture

DESCRIPTION (provided by applicant) Mitochondria play a vital role in cell physiology and the response to environmental stress. A number of cellular toxins, including rotenone, 1-methyl-4-phenylpyridine (MPP+) and paraquat act to impair mitochondrial electron transport, generating ROS. Reactive oxygen species are also important in the toxicity of arsenic, amyloid, and ceramide. Mitochondria have been viewed as a potential source of ROS that may contribute to Parkinson's disease, aging and other pathological conditions. Since mitochondria contain only a small 16.5 kb mtDNA genome, encoding only 13 proteins, the organelle depends on the nucleus for most gene products, including all of the factors required for DNA replication, expression and repair. Recent studies from our laboratory and others have revealed an increasing collection of proteins that function in both mitochondria and other cellular compartments. A number of these proteins function in repair of oxidative damage to mtDNA. One significant consequence of mitochondrial pathology is the generation of mutations in mtDNA, many of which have a tissue-specific incidence, occurring most commonly in postreplicative tissues such as nerve and muscle. The investigators propose to test the hypothesis that mitochondria in differentiated cells may contain a different complement of proteins than actively dividing cells which may predispose post-replicative cells to a higher rate of mtDNA mutations or may alter the ability of cells to enter apoptosis. To accomplish this, they will study the effect of oxidative stress on the mitochondrial proteome in both embryonal carcinoma cells that are actively dividing and in cells that have been induced to differentiate along a neuronal pathway. Both 2-D gel methods and quantitative isotope-coded affinity tag (ICAT) methods will be used to compare the abundance of mitochondrial proteins in control cells and cells exposed to oxidative stress. The broad proteomic screen will permit the discovery of novel gene products not previously known to function in mitochondria. Data will be analyzed to provide new insights into networks of proteins acting to repair oxidative damage to mtDNA or to detoxify ROS.

描述（由申请人提供） 线粒体在细胞生理和对环境压力的反应中起着至关重要的作用。 许多细胞毒素，包括烤面包酮，1-甲基-4-苯基吡啶（MPP+）和Paraquat ACT，以损害线粒体电子转运，产生ROS。 活性氧在砷，淀粉样蛋白酶和神经酰胺的毒性中也很重要。 线粒体被视为可能导致帕金森氏病，衰老和其他病理状况的潜在ROS来源。 由于线粒体仅包含一个小的16.5 kb mtDNA基因组，仅编码13种蛋白质，因此细胞器取决于大多数基因产物的细胞核，包括DNA复制，表达和修复所需的所有因素。 我们的实验室和其他人的最新研究表明，在线粒体和其他细胞室中起作用的蛋白质越来越多。 这些蛋白质的许多在修复对mtDNA的氧化损伤方面起作用。 线粒体病理学的一个重要结果是MTDNA的突变产生，其中许多具有组织特异性的发病率，最常见于神经和肌肉等上皮组织中。 研究者建议检验以下假设：分化细胞中的线粒体可能包含与主动分裂细胞不同的蛋白质补体，这些细胞可能会使复制后细胞倾向于更高的mtDNA突变发生率，或者可能会改变细胞的能力。 为此，他们将研究氧化应激对两个胚胎癌细胞中线粒体蛋白质组的影响，这些细胞正在积极分裂以及已诱导沿神经元途径区分的细胞中。 二维凝胶方法和定量同位素编码的亲和力标签（ICAT）方法将用于比较对照细胞和暴露于氧化应激的细胞中线粒体蛋白的丰度。 广泛的蛋白质组学筛选将允许发现以前未知在线粒体中起作用的新基因产物。 数据将进行分析，以提供对作用于修复MTDNA氧化损伤或排毒ROS的蛋白质网络的新见解。