S-Nitrosation in Nitric Oxide Signaling

一氧化氮信号传导中的 S-亚硝化

基本信息

批准号：
7458666
负责人：
WILLIAM R. MONTFORT
金额：
$ 27.05万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7458666
关键词：
Alcohol dehydrogenase Allosteric Regulation Asthma Binding Biochemical Biochemistry Biological Models Biology Blood Pressure Cancer cell line Cardiovascular Diseases Catalysis Cell Line Cells Chemicals Chemistry Cyclic GMP Development Diabetes Mellitus Disease Electrons Engineering Event Genes Glutathione Glutathione Metabolism Pathway Glutathione Reductase Heart Diseases Heme Human In Vitro Insecta Investigation Link Malignant Neoplasms Memory Metabolism Modeling Modification Molecular Genetics Muscle relaxation phase NADH Nitric Oxide Nitric Oxide Signaling Pathway Nitrosation Oxidoreductase Physiological Play Positioning Attribute Production Protein Conformation Proteins Reaction Recombinants Regulation Resolution Role SKIL gene Signal Pathway Signal Transduction Soluble Guanylate Cyclase Structure Sulfhydryl Compounds System Tetracycline Tetracyclines Therapeutic Agents Therapeutic Uses Thinking Thioredoxin base cell growth regulation design drug discovery human NOS2A protein human TXN protein in vivo inhibitor/antagonist insight programs research study structural biology tool

项目摘要

DESCRIPTION (provided by applicant): Protein S-nitrosation occurs throughout biology and plays a regulatory role in all major diseases, including cardiovascular disease, diabetes, asthma and cancer. Yet, the chemical and structural biology underlying these events remains undiscovered. We will bridge this gap through crystallographic, biochemical and molecular genetics investigations of three key proteins: thioredoxin, soluble guanylate cyclase and glutathione S-nitroso reductase (GSNOR). Together, these proteins perform much of the RSNO-based chemistry thought to occur in the cell and will allow us to investigate the structure and dynamics of these activities in normal and disease states. Specific Aim 1: Transnitrosation reactions involving human thioredoxin. Thioredoxin has emerged as a key intermediate in protein transnitrosation and is also implicated in several diseases, including cancer and heart disease. We have achieved a crystal structure of S-nitrosated human thioredoxin through transnitrosation by S-nitrosogluatathione (GSNO), the first such structure to be described. We are now in a position to explore the structure and biochemistry of this modification in vitro and in a model cancer cell line. Specific Aim 2: GSNO stimulation of soluble guanylate cyclase. The best understood pathway for nitric oxide signaling is through binding to heme in soluble guanylate cyclase (sGC), which initiates cGMP production to regulate physiological activities such as blood pressure and memory formation. We have discovered that GSNO is also a potent stimulant of sGC. We will undertake biochemical and molecular genetic experiments to reveal the role of GSNO in sGC function Specific Aim 3: GSNO metabolism by GSNO reductase. Much of the nitric oxide produced in vivo is initially captured by glutathione as GSNO, which participates in SNO chemistry and is metabolized by GSNOR. We have determined a high resolution structure for GSNOR and discovered an inhibitor of the protein. We now propose a program in drug discovery and biochemistry targeted to GSNOR.

描述（由申请人提供）：蛋白质S-硝化发生在整个生物学过程中，并在所有主要疾病（包括心血管疾病，糖尿病，哮喘和癌症）中发挥调节作用。然而，这些事件的基础化学和结构生物学仍然没有发现。我们将通过三种关键蛋白的晶体学，生化和分子遗传学研究来弥合这一差距：硫氧还蛋白，可溶性鸟苷酸环化酶和谷胱甘肽S-硝基还原酶（GSNOR）。总之，这些蛋白质执行了大部分基于RSNO的化学物质，认为这些化学物质会发生在细胞中，并使我们能够研究正常和疾病状态中这些活动的结构和动力学。特定目的1：涉及人硫氧还蛋白的转硝化反应。硫氧还蛋白已成为蛋白质转屈的关键中间体，还与多种疾病有关，包括癌症和心脏病。我们通过S-硝基氟硫代（GSNO）的转硝化（GSNO）实现了S-硝化的人硫蛋白的晶体结构，这是第一个要描述的结构。现在，我们可以在体外和模型癌细胞系中探索这种修饰的结构和生物化学。特定目标2：GSNO刺激可溶性鸟烯酸酸酯环化酶。一氧化氮信号传导的最佳理解途径是通过在可溶性鸟苷酸环化酶（SGC）中与血红素结合，该鸟烯醇环酶（SGC）启动CGMP产生以调节生理活性，例如血压和记忆形成。我们发现GSNO也是SGC的有效刺激物。我们将进行生化和分子遗传实验，以揭示GSNO在SGC功能特定目标中的作用3：GSNO还原酶的GSNO代谢。在体内产生的一氧化氮最初被谷胱甘肽捕获为GSNO，它参与SNO化学，并被GSNOR代谢。我们已经确定了GSNOR的高分辨率结构，并发现了蛋白质的抑制剂。现在，我们建议针对GSNOR的药物发现和生物化学计划。