Chemical approaches for detecting S-nitrosothiols

检测 S-亚硝基硫醇的化学方法

• 批准号: 7887896
• 负责人: Ming Xian
• 金额: $ 25.54万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7887896
• 关键词: Acetylation; Alkynes; Biological; Biomedical Research; Biotin; Cells; Chemicals; Chemistry; Collaborations; Complex; Cysteine; Data; Detection; Disulfides; Functional disorder; Goals; Label; Lead; Ligation; Link; Mediating; Methods; Modeling; Modification; Nitric Oxide; Nitrosation; Phosphines; Phosphorylation; Physiological; Physiology; Play; Polymers; Post-Translational Protein Processing; Process; Property; Proteins; Proteomics; Reaction; Reagent; Reporter; Research; Research Personnel; Role; SKIL gene; Sampling; Scheme; Series; Signal Transduction; Signaling Molecule; Solid; System; Techniques; Testing; Tissues; Vision; Work; base; biological systems; complex biological systems; dehydroalanine; design; insight; interest; public health relevance; research study; sulfenamide; ward

DESCRIPTION (provided by applicant): Nitric oxide is a cell-signaling molecule involved in a number of physiological and pathophysiological processes. Modification of cysteine residues by nitric oxide, i.e. S- nitrosation, changes the function of a broad spectrum of proteins. This reaction represents an important post-translational modification that transduces nitric oxide- dependent signals. However, the detection and quantification of S-nitrosation in biological samples remains a challenge because of the lability of the S-nitrosation products: S-nitrosothiols (RSNOs). Our group recently developed a series of new reactions for RSNOs, which include bis-ligation, reductive ligation, and reductive elimination (to form dehydroalanine). These reactions selectively target on RSNOs and convert them to stable/detectable products. Based on control experiments, we hypothesize these reactions can be used to design new methods for the detection of RSNOs. In this project, we plan to pursue the following four Specific Aims: (1) to study bis-ligation based methods for detecting protein S-nitrosation; (2) to study reductive ligation based methods for detecting protein S-nitrosation; (3) to study dehydroalanine formation from S-nitrosocysteines and the applications in the detection of protein S- nitrosation; and (4) to validate the applications of new RSNO detection methods in biological systems. These studies will advance the understanding of the chemical and biological properties of RSNOs and permit studies of S-nitrosation-related nitric oxide signal transducation in complex systems. PUBLIC HEALTH RELEVANCE: S-Nitrosation of protein cysteine residues plays important roles in physiology and pathophysiology, while the detection of S-nitrosation is still a challenge. The research proposed here will provide new insights into the chemistry of S-nitrosation and lead to new techniques for the detection of S-nitrosation in biological samples, which should have great impact on biomedical research.

描述（由申请人提供）：一氧化氮是一种参与许多生理和病理生理过程的细胞信号分子。一氧化氮对半胱氨酸残基的修饰，即 S-亚硝化，改变了多种蛋白质的功能。该反应代表了一种重要的翻译后修饰，可转导一氧化氮依赖性信号。然而，由于 S-亚硝化产物：S-亚硝基硫醇 (RSNO) 的不稳定，生物样品中 S-亚硝化的检测和定量仍然是一个挑战。我们课题组最近开发了一系列针对 RSNO 的新反应，包括双连接、还原连接和还原消除（形成脱氢丙氨酸）。这些反应选择性地针对 RSNO 并将其转化为稳定/可检测的产物。基于对照实验，我们假设这些反应可用于设计检测 RSNO 的新方法。在本项目中，我们计划实现以下四个具体目标：（1）研究基于双连接的蛋白质S-亚硝化检测方法； (2) 研究基于还原连接的蛋白质S-亚硝化检测方法； (3) 研究S-亚硝基半胱氨酸形成脱氢丙氨酸及其在蛋白质S-亚硝化检测中的应用； (4)验证新的RSNO检测方法在生物系统中的应用。这些研究将增进对 RSNO 化学和生物特性的理解，并允许研究复杂系统中与 S-亚硝化相关的一氧化氮信号转导。 公共健康相关性：蛋白质半胱氨酸残基的 S-亚硝化在生理学和病理生理学中发挥着重要作用，而 S-亚硝化的检测仍然是一个挑战。这里提出的研究将为S-亚硝化化学提供新的见解，并带来检测生物样品中S-亚硝化的新技术，这将对生物医学研究产生重大影响。