Oxidative stress, thiols, and redox profiles.
Oxidative stress, thiols, and redox profiles.
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DOI：
10.1007/978-1-61779-867-2_21
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Hansen, Jason M
中科院分区：
文献类型：
--
作者：
Harris, Craig;Hansen, Jason M
氧化应激已被认为是导致大量发育毒物毒性的一个因素。氧化应激的传统定义是，细胞内还原生物分子和氧化生物分子之间的平衡发生变化，有利于后者，导致有害于重要细胞功能的变化，最终可能导致畸形和死亡。谷胱甘肽(GSH)/谷胱甘肽二硫化物(GSSG)氧化还原对因其浓度高、直接作为抗氧化剂和细胞保护剂而成为表征氧化应激的传统标记物。GSH通过结合、氧化或输出的稳态耗竭经常被报道为引发氧化应激的唯一标准和无数相关的有害后果。许多其他的，大多是定性的，也有报道表明氧化应激已经发生，但并不总是清楚它们在多大程度上反映了细胞的状态或其功能。我们对氧化还原信号和活性氧簇(ROS)、硫醇、氧化剂分子和细胞抗氧化剂的作用的新理解，都作为第二信使，促使我们根据真正的翻译后蛋白质硫醇修饰的变化重新定义氧化应激，这些修饰是氧化还原调控的核心。基于硫醇的氧化还原对，如GSH/GSSG、半胱氨酸/半胱氨酸(Cys/Cyss)、硫氧还蛋白还原/硫氧还蛋白氧化(Trx(Red)/Trx(Ox))形成独立的信号节点，选择性地调节发育事件，并与细胞内氧化还原电位的变化密切相关。在发育过程中，准确评估自由基增加的后果最好使用一系列测量方法，包括细胞内氧化还原电位、ROS、生物分子的氧化还原状态以及特定氧化还原信号节点的诱导变化。提出了测定ROS产生、可溶性硫醇氧化、氧化还原电位的方法，以及评价特定蛋白质硫醇氧化状态的蛋白质组学方法。
Oxidative stress has been recognized as a contributing factor in the toxicity of a large number of developmental toxicants. Traditional definitions of oxidative stress state that a shift in the balance between reduced and oxidized biomolecules within cells, in favor of the latter, result in changes that are deleterious to vital cell functions and can culminate in malformations and death. The glutathione (GSH)/glutathione disulfide (GSSG) redox couple has been the traditional marker of choice for characterization of oxidative stress because of its high concentrations and direct roles as antioxidant and cellular protectant. Steady state depletion of GSH through conjugation, oxidation, or export has often been reported as the sole criteria for invoking oxidative stress and a myriad of associated deleterious consequences. Numerous other, mostly qualitative, observations have also been reported to suggest oxidative stress has occurred but it is not always clear how well they reflect the state of a cell or its functions. Our emerging understanding of redox signaling and the roles of reactive oxygen species (ROS), thiols, oxidant molecules, and cellular antioxidants, all acting as second messengers, has prompted a redefinition of oxidative stress based on changes in the real posttranslational protein thiol modifications that are central to redox regulation and control. Thiol-based redox couples such as GSH/GSSG, cysteine/cystine (cys/cySS), thioredoxin-reduced/thioredoxin-oxidized (TRX(red)/TRX(ox)) form independent signaling nodes that selectively regulate developmental events and are closely linked to changes in intracellular redox potentials. Accurate assessment of the consequences of increased free radicals in developing conceptuses should best be made using a battery of measurements including the quantitative assessment of intracellular redox potential, ROS, redox status of biomolecules, and induced changes in specific redox signaling nodes. Methods are presented for a determination of ROS production, soluble thiol oxidation, redox potential, and a proteomic approach to evaluate the thiol oxidation state of specific proteins.