NO signaling by a Soluble Guanylyl Cyclase-Thioredoxin transnitrosation complex

可溶性鸟苷酸环化酶-硫氧还蛋白转亚硝基复合物的 NO 信号转导

基本信息

批准号：
8894270
负责人：
ANNIE V BEUVE
金额：
$ 41.16万
依托单位：
RBHS-NEW JERSEY MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8894270
关键词：
Angiotensin II Apoptosis Apoptosis Regulator Apoptotic Binding Biochemical Bioinformatics Biological Biological Assay Cardiac Cardiac Myocytes Cardiovascular Physiology Cardiovascular system Cell Line Cell Nucleus Cell Survival Cells Cessation of life Complex Consensus Sequence Cyclic GMP Cysteine Data Environment Functional disorder Guanosine Triphosphate Heart Heart Hypertrophy Heart failure In Vitro Investigation Lead Mass Spectrum Analysis Mediating Mediator of activation protein Modeling Modification Mus Mutagenesis Myocardium Nitric Oxide Nitrosation Nuclear Translocation Oxidation-Reduction Pathway interactions Play Post-Translational Protein Processing Process Property Proteins Proteome Proteomics Regulation Role SKIL gene Signal Pathway Signal Transduction Signaling Molecule Site Smooth Muscle Myocytes Soluble Guanylate Cyclase Specificity Stress Sulfhydryl Compounds Thioredoxin Transgenic Mice angiogenesis base cGMP production heme a human CLIC4 protein mutant nitrosative stress novel oxidation prevent protein function protein protein interaction public health relevance research study response small hairpin RNA

项目摘要

DESCRIPTION (provided by applicant): Nitric oxide (NO) is an important signaling molecule that regulates diverse functions relevant to cardiovascular function, apoptosis and angiogenesis. NO is best known for its ability to stimulate soluble guanylyl cyclase (sGC) to produce cGMP and stimulate its downstream signaling pathways. However, NO can also covalently modify cysteines via S-nitrosation (addition of a NO moiety to the cysteine of a protein, SNO). Although this reversible post-translational modification is increasingly recognized as an important regulatory mechanism of protein function, and to play a role in cardiac protection, dynamic regulation of protein nitrosation specificity is poorly understood. Our collaborative team has made the exciting observation that sGC, the key NO receptor, modulates the level of nitrosation of specific proteins in cardiomyocytes and smooth muscle cells. Preliminary data showed that sGC increases nitrosation by a protein-protein interaction-driven SNO transfer (transnitrosation). Moreover, this increased nitrosation is due, for a specific subset of proteins, to the association of sGC with thioredoxin 1 (Trx1), a cardiac protective thiol-redox protein with both transnitrosation and denitrosation activities. Initial mass spectrometry and biochemical analyses showed that sGC transnitrosates Trx1, which in turn nitrosates a specific subset of targets, a finding supported by shTrx1 knockdown experiments in cardiomyocytes. These novel observations lead to the provocative idea that sGC modulates S-nitrosation specificity via a transnitrosation cascade that includes an S-nitrosated Trx1 intermediate. This study aims to answer three critical questions based on this hypothesis. Aim1: What is the mechanism of sGC transnitrosation of Trx1? We will identify key cysteines (Cys) responsible for sGC transfer of SNO to Trx1 and for interaction via mutagenesis and biochemical analyses. Aim2: What are the specific targets of the sGC/Trx1 transnitrosation cascade and the mechanisms underlying target specificity? Using novel and highly specific proteomics approaches, we will quantify the SNO-proteomes modulated by the sGC/Trx1 transnitrosation cascade under nitrosative and oxidative conditions and determine consensus sequence motifs among the target proteins. Aim3: Is sGC-mediated transnitrosation an anti-apoptotic mechanism? NO signaling and Trx1 are crucial components of the anti-apoptotic response to stress. Among the sGC/Trx1 transnitrosation targets identified is the chloride intracellular channel 4 (CLIC4), a regulator of apoptosis, whose nuclear translocation is modulated via specific nitrosation. We will determine whether sGC/Trx1 transnitrosation of CLIC4 is an important inhibitory mechanism of angiotensin II-induced apoptosis of cardiomyocytes, underlying the potential role of this newly discovered transnitrosation cascade in cardiac remodeling following heart failure. This multi-PI project could lead to the discovery of novel cardioprotective pathway driven by specific S-nitrosation.

描述（由申请人证明）：一氧化氮（NO）是一个重要的信号，可以调节与心血管功能，凋亡和血管生成有关的多种功能。在SNO）。这种可逆的翻译后修饰是蛋白质功能的重要调节机制，并且在心脏保护中发挥作用，对蛋白质硝化规范的动态调节使得很差。关键的受体，心肌细胞和肌肉细胞中特异性蛋白的硝化水平通过蛋白质驱动的SNO转移（Transnitrosation）提高了硝化作用。硫氧还蛋白1（TRX1）具有两种跨硝化活性的心脏保护性硫醇 - 雷多斯蛋白。这项研究旨在解决三个Cree creesthothothesis。 SGC/TRX1反元素级联的特定目标是使用新颖和高度特异性的蛋白质组学方法的基础机制，我们将量化SNO-Proteomes模块化的模块化并确定至关重要的。对应激的抗凋亡反应的组成部分（CLIC4），凋亡的调节剂核易位是通过特异性的硝化来调节的。导致发现由特定S-硝化驱动的新型心脏保护途径。