Sheehan David
Department of Biochemistry, University College Cork, Lee Maltings, Prospect Row, Mardyke, Cork, Ireland.
Biochem Biophys Res Commun. 2006 Oct 20;349(2):455-62. doi: 10.1016/j.bbrc.2006.08.124. Epub 2006 Aug 31.
Oxidative stress arises when cellular defenses against molecular oxygen and its by-products (reactive oxygen species; ROS) are overcome leading to covalent modification of lipids, DNA, and protein. Redox-based modification of proteins can be conveniently studied by proteomic analysis using two-dimensional electrophoresis (2D SDS-PAGE). Despite some technical shortcomings, this technique allows rapid and quantitative analysis of paired samples, the visualization of discrete protein spots, and provides a robust platform for subsequent analysis and identification of specific proteins. Exposure to oxidative stress introduces a wide range of reversible or irreversible alterations to amino acid side chains. These include carbonylation, glutathionylation, formation of mixed disulphides, effects on disulphide bridge patterns, ubiquitinylation, and racemization. Identification of proteins targeted for specific modification adds a deeper dimension to the dissection of effects of oxidative stress on the proteome with potentially far-reaching implications. This article describes key methodologies now available for identification of redox-based modifications in proteins separated by 2D SDS-PAGE.
当细胞对分子氧及其副产物（活性氧；ROS）的防御机制被打破，导致脂质、DNA和蛋白质发生共价修饰时，就会产生氧化应激。基于氧化还原的蛋白质修饰可以通过二维电泳（2D SDS-PAGE）的蛋白质组学分析方便地进行研究。尽管存在一些技术缺陷，但该技术允许对配对样本进行快速定量分析，可视化离散的蛋白质斑点，并为后续特定蛋白质的分析和鉴定提供了一个强大的平台。暴露于氧化应激会使氨基酸侧链发生广泛的可逆或不可逆变化。这些变化包括羰基化、谷胱甘肽化、混合二硫键的形成、对二硫键模式的影响、泛素化和消旋化。鉴定针对特定修饰的蛋白质为剖析氧化应激对蛋白质组的影响增添了更深层次的维度，可能具有深远的意义。本文介绍了目前可用于鉴定通过2D SDS-PAGE分离的蛋白质中基于氧化还原修饰的关键方法。
