Regulation of Soluble Guanylyl Cyclase, the NO-Receptor

可溶性鸟苷酸环化酶（NO 受体）的调节

• 批准号: 8452113
• 负责人: ANNIE V BEUVE
• 金额: $ 8.28万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8452113
• 关键词: Affect; Affinity; American; Angiotensin II; Animal Model; Atherosclerosis; Binding; Biochemical; Biological Availability; Biological Process; Biology; Blood Vessels; Cardiovascular Diseases; Cardiovascular Pathology; Cardiovascular system; Catalytic Domain; Complex; Cyclic GMP; Cysteine; Data; Development; Dimerization; Dominant-Negative Mutation; Endothelium-Dependent Relaxing Factors; Enzymes; Erectile dysfunction; Etiology; Functional disorder; Funding; Gases; Goals; Guanosine Triphosphate; Heart Hypertrophy; Heme; Homeostasis; Hypertension; Impairment; Infection; Kinetics; Link; Maps; Modeling; Modification; Molecular; Mutate; Neurons; Nitrates; Nitric Oxide; Nitrosation; Oxidation-Reduction; Oxidative Stress; Pathology; Pathway interactions; Physiological; Physiology; Production; Protein Disulfide Isomerase; Proteins; Proteomics; Reaction; Regulation; Relaxation; Reperfusion Injury; Resistance; Role; Signal Transduction; Smooth Muscle Myocytes; Soluble Guanylate Cyclase; Structure; Sulfhydryl Compounds; Synaptic plasticity; System; Testing; Thioredoxin; arteriole; base; clinically relevant; desensitization; disulfide bond; experience; heme a; in vivo; inhibitor/antagonist; innovation; molecular dynamics; mutant; nitrosative stress; overexpression; receptor; response; small molecule

DESCRIPTION (provided by applicant): Since the discovery that the endothelium derived relaxing factor (EDRF) was the endogenous toxic gas nitric oxide (NO), an astonishing number of physiological functions have been attributed to NO. Despite the widely recognized importance of NO, little is known about the mechanism of regulation of the NO receptor, the soluble guanylyl cyclase (sGC). sGC is a heme containing heterodimer that catalyzes the formation of cGMP from the substrate GTP. Upon binding of NO, the sGC is activated several hundred fold. The sGC is a multi-domain signaling enzyme that contains the NO receptor-heme domain, a dimerization domain and the effector-catalytic domain. It is not known how the NO signal is propagated to the catalytic domain of sGC. We recently discovered that sGC is desensitized by S-nitrosylation, the addition of a NO moiety to the free thiol of a specific cysteine (Cys). There s now substantial evidence that thiol redox reactions are critical and dynamic regulators of sGC function and that thiol modifications of sGC have clinical relevance as they are associated with decreased vascular reactivity in hypertension. However, the mechanisms by which modification of reactive Cys modulate sGC activation, NO signal transduction to the catalytic domain, sGC basal activity, domain interactions and NO-heme affinity have yet to be explored. The proposed studies seek to understand the structural and molecular basis of mechanisms of regulation of the sGC and how disruption of these mechanisms contributes to development of cardiovascular pathologies. We will use purified enzymes, cellular system and animals models to conduct structure-function studies, molecular dynamics simulation, biochemical and kinetics analysis and applied physiology for the following aims: 1) define the role of thiol Cys in the molecular mechanism of activation of sGC; 2) investigate the modulation of sGC by the thiol-reducing protein thioredoxin and 3) establish how thiol-dependent dysfunction of sGC contributes to hypertension and cardiac hypertrophy. Understanding the mechanisms of regulation of sGC will be key to uncovering the molecular basis of some types of hypertension, atherosclerosis and erectile dysfunction, which affect more than 60 millions Americans.

描述（由申请人提供）：由于发现衍生的松弛因子（EDRF）的发现是内源性有毒气体一氧化氮（NO），因此已经将大量生理功能归因于NO。尽管NO具有广泛认可的重要性，但对NO受体调节的机理，可溶性鸟叶基环酶（SGC）知之甚少。 SGC是一种含有异二聚体的血红素，可催化从底物GTP形成CGMP。 NO结合后，将SGC激活数百倍。 SGC是一种多域信号传导酶，其中包含NO受体 - 血红素结构域，二聚域和效应催化域。尚不清楚NO信号如何传播到SGC的催化结构域。我们最近发现，S-in硝基化脱敏，在特定半胱氨酸（CYS）的游离硫醇（CYS）的游离硫醇中添加了无部分。现在有大量证据表明，硫醇氧化还原反应是SGC功能的关键和动态调节剂，并且SGC的硫醇修饰具有临床相关性，因为它们与高血压血管反应的降低相关。但是，反应性Cys修饰调节SGC激活的机制，尚未探索催化域，SGC基础活性，域相互作用和无血糖亲和力的信号转导。拟议的研究旨在了解SGC调节机制的结构和分子基础，以及这些机制的破坏如何有助于心血管病理的发展。我们将使用纯化的酶，细胞系统和动物模型进行结构功能研究，分子动力学模拟，生化和动力学分析以及应用生理学的以下目的：1）定义硫醇Cys在SGC激活分子机制中的作用； 2）研究通过还原硫醇蛋白硫氧还蛋白对SGC的调节，3）确定SGC的硫醇依赖性功能障碍如何有助于高血压和心脏肥大。了解SGC调节的机制将是揭示某些类型的高血压，动脉粥样硬化和勃起功能障碍的分子基础的关键，这会影响超过6000万美国人。