Mass Spectrometry And Oxidative Stress

质谱法和氧化应激

• 批准号: 6507359
• 负责人: Kenneth Tomer
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6507359
• 关键词: DNA damage; adduct; aging; cytochrome c; electron spin resonance spectroscopy; free radicals; glutathione; horses; hydrogen peroxide; inflammation; lipid peroxides; mass spectrometry; matrix assisted laser desorption ionization; mitochondria; nitroso compounds; oxidation reduction reaction; oxidative stress; peroxidation; phosphorylation; protein degradation

Summary of Work: The study of oxidative stress has attracted considerable interest and has been the focus of much research in recent years. Cumulative oxidative damage to tissues has been implicated in a number of disease states, e.g. the aging process, cancer, and ischemia reperfusion. The study of oxidative stress in the mitochondria has shown that hydrogen peroxide is produced via the incomplete reduction of oxygen during oxidative phosphorylation. Hydrogen peroxide levels are kept relatively low under normal physiological conditions. Under certain conditions, such as inflammation, excessive amounts of hydrogen peroxide are produced. The production of excess hydrogen peroxide is thought to precede several occurrences, such as lipid peroxidation, DNA and/or protein damage, and glutathione depletion, that are characteristic of oxidative stress. In a long-term collaboration with the epr group (Mason/LPC) we have been investigating the sites of radical formation on proteins and peptides and have been determining the specific site on the amino acid which undergoes radical formation during oxidation by determining the structure of the amino acid spin-trap covalent adduct. This information is important to our understanding of the damage that can occur by free-radical oxidation of proteins and how that damage occurs. Investigation of the mechanism of development of Parkinson's disease: In collaboration with R. Mason (LPC) and G. Pielak (UNC), we are involved in a project whose aims is to probe how a-synuclein undergoes Lewy body formation. The working hypothesis is that oxidative damage to heme proteins such as cyt c leads to release of the cyt c from the mitochondria. Upon release, the cyt c free radical can react further with other proteins, such as a-synuclein. These secondary free-radical proteins can then undergo either chemical cross-linking or conformational changes that lead to aggregation.

工作摘要：氧化应激的研究引起了人们的极大兴趣，近年来一直是许多研究的重点。对组织的累积氧化损害与许多疾病状态有关，例如衰老过程，癌症和缺血再灌注。线粒体中氧化应激的研究表明，过氧化氢是通过氧化磷酸化过程中氧气不完全减少而产生的。在正常的生理条件下，过氧化氢水平保持相对较低。在某些条件下，例如炎症，产生了过多的过氧化氢。过多的过氧化氢的产生被认为是在几次发生之前的生产，例如脂质过氧化，DNA和/或蛋白质损伤以及谷胱甘肽耗竭，这些损害是氧化应激的特征。 在与EPR组（MASON/LPC）的长期合作中，我们一直在研究蛋白质和肽对根部形成的部位，并一直在确定氨基酸上的特定位点，该氨基酸通过确定氨基酸自旋 - 触发型物质的结构而在氧化过程中进行了自由基形成。 这些信息对于我们对蛋白质的自由基氧化可能发生的损害以及该损害如何发生的损害很重要。 调查帕金森氏病发展机制：与R. Mason（LPC）和G. Pielak（UNC）合作，我们参与了一个项目，其目的是探究A-突触核蛋白如何经历Lewy的身体形成。工作假设是，对血红素蛋白（例如Cyt c）的氧化损伤导致细胞体从线粒体释放。释放后，Cyt C自由基可以与其他蛋白质（例如A-核蛋白）进一步反应。然后，这些二次自由基蛋白可以进行化学交联或构象变化，从而导致聚集。