Chemical approaches for detecting S-nitrosothiols

检测 S-亚硝基硫醇的化学方法

• 批准号: 8727585
• 负责人: Ming Xian
• 金额: $ 27.93万
• 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 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8727585
• 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; Protein S; Proteins; Proteomics; Reaction; Reagent; Reporter; Research; Research Personnel; Role; S-Nitrosothiols; SKIL gene; Sampling; Scheme; Series; Signal Transduction; Signaling Molecule; Solid; System; Techniques; Testing; Tissue Sample; Tissues; Vision; Work; base; biological systems; complex biological systems; dehydroalanine; design; insight; interest; public health relevance; research study; sulfenamide; ward; 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; Protein S; Proteins; Proteomics; Reaction; Reagent; Reporter; Research; Research Personnel; Role; S-Nitrosothiols; SKIL gene; Sampling; Scheme; Series; Signal Transduction; Signaling Molecule; Solid; System; Techniques; Testing; Tissue Sample; 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.

描述（由申请人提供）：一氧化氮是参与许多生理和病理生理过程的细胞信号分子。一氧化氮（即S-硝化）修饰半胱氨酸残基改变了蛋白质广谱的功能。该反应代表了一个重要的翻译后修饰，该修饰可转导一氧化氮依赖性信号。但是，由于S-硝化产物的不稳定：S-硝基硫醇（RSNOS），生物样品中S-硝化的检测和定量仍然是一个挑战。我们的小组最近为RSNO开发了一系列新反应，其中包括双 - 连接，还原结扎和还原性消除（形成脱氢丙氨酸）。这些反应选择性地靶向RSNO，并将其转换为稳定/可检测的产品。基于控制实验，我们假设这些反应可用于设计用于检测RSNO的新方法。在该项目中，我们计划追求以下四个特定目的：（1）研究基于双 - 结构的方法检测蛋白质S-硝化的方法； （2）研究基于还原连接的方法检测蛋白质S-硝化的方法； （3）研究从S-硝基糖粉和检测蛋白质S-硝基化的应用中研究脱氢丙氨酸的形成； （4）验证新的RSNO检测方法在生物系统中的应用。这些研究将提高对RSNO的化学和生物学特性的了解，并允许研究复杂系统中与S-硝化相关的一氧化氮信号转导。