Mitochondrial Response to Oxidative Stress

线粒体对氧化应激的反应

基本信息

批准号：
7103696
负责人：
DANIEL F. BOGENHAGEN
金额：
$ 36.74万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-09-10 至 2008-07-31
项目状态：
已结题

项目摘要

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）线粒体在细胞生理学和对环境压力的反应中发挥着至关重要的作用。许多细胞毒素，包括鱼藤酮、1-甲基-4-苯基吡啶 (MPP+) 和百草枯，都会损害线粒体电子传输，产生 ROS。活性氧在砷、淀粉样蛋白和神经酰胺的毒性中也很重要。线粒体被认为是活性氧的潜在来源，可能导致帕金森病、衰老和其他病理状况。由于线粒体仅含有 16.5 kb 的 mtDNA 基因组，仅编码 13 种蛋白质，因此细胞器依赖于细胞核来产生大多数基因产物，包括 DNA 复制、表达和修复所需的所有因子。我们实验室和其他实验室的最新研究表明，在线粒体和其他细胞区室中发挥作用的蛋白质越来越多。其中许多蛋白质具有修复线粒体 DNA 氧化损伤的功能。线粒体病理学的一个重要后果是线粒体 DNA 突变的产生，其中许多突变具有组织特异性，最常见于复制后组织，如神经和肌肉。研究人员提议检验这样的假设：分化细胞中的线粒体可能含有与活跃分裂细胞不同的蛋白质补体，这可能会使复制后细胞发生更高率的 mtDNA 突变，或者可能改变细胞进入凋亡的能力。为了实现这一目标，他们将研究氧化应激对活跃分裂的胚胎癌细胞和沿神经元通路诱导分化的细胞中线粒体蛋白质组的影响。二维凝胶方法和定量同位素编码亲和标签 (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的在线粒体中发挥作用的新基因产物。将对数据进行分析，以提供对修复 mtDNA 氧化损伤或解毒 ROS 的蛋白质网络的新见解。