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Abstract：
Reactive nitrogen species, such as nitric oxide (NO), exert their biological activity in large part through post-translational modification of cysteine residues, forming S-nitrosothiols. This chemical reaction proceeds via a process that we and our colleagues have termed protein S-nitrosylation. Under conditions of normal NO production, S-nitrosylation regulates the activity of many normal proteins. However, in degenerative conditions characterized by nitrosative stress, increased levels of NO lead to aberrant S-nitrosylation that contributes to the pathology of the disease. Thus, S-nitrosylation has been implicated in a wide range of cellular mechanisms, including mitochondrial function, proteostasis, transcriptional regulation, synaptic activity, and cell survival. In recent years, the research area of protein S-nitrosylation has become prominent due to improvements in the detection systems as well as the demonstration that protein S-nitrosylation plays a critical role in the pathogenesis of neurodegenerative and other neurological disorders. To further promote our understanding of how protein S-nitrosylation affects cellular systems, guidelines for the design and conduct of research on S-nitrosylated (or SNO-)proteins would be highly desirable, especially for those newly entering the field. In this review article, we provide a strategic overview of designing experimental approaches to study protein S-nitrosylation. We specifically focus on methods that can provide critical data to demonstrate that an S-nitrosylated protein plays a (patho-)physiologically-relevant role in a biological process. Hence, the implementation of the approaches described herein will contribute to further advancement of the study of S-nitrosylated proteins, not only in neuroscience but also in other research fields.
摘要：
活性氮物种，如一氧化氮（NO），在很大程度上通过半胱氨酸残基的翻译后修饰发挥其生物活性，形成S-亚硝基硫醇。该化学反应通过我们和我们的同事称为蛋白质S-亚硝基化的过程进行。在正常NO产生的条件下，S-亚硝基化调节许多正常蛋白质的活性。然而,在以亚硝基应激为特征的退行性疾病中，NO水平的增加导致导致疾病病理的异常S-亚硝基化。因此，S-亚硝基化涉及广泛的细胞机制，包括线粒体功能,proteostasis,转录调控，突触活动，细胞存活。近年来,由于检测系统的改进以及蛋白质S-亚硝基化在神经退行性疾病和其他神经系统疾病的发病机理中起关键作用的证明，蛋白质S-亚硝基化的研究领域变得突出。为了进一步促进我们对蛋白质S-亚硝基化如何影响细胞系统的理解，设计和进行S-亚硝基化（或SNO-）蛋白质研究的指南将是非常可取的，尤其是那些刚进入这个领域的人。在这篇评论文章中，我们提供了设计实验方法来研究蛋白质S-亚硝基化的战略概述。我们特别关注可以提供关键数据的方法，以证明S-亚硝基化蛋白质在生物学过程中起（病理）生理相关作用。因此，本文所述方法的实施将有助于进一步推进S-亚硝基化蛋白的研究，不仅在神经科学领域，而且在其他研究领域。
