Activation Mechanism of Soluble Guanylate Cyclase

可溶性鸟苷酸环化酶的激活机制

• 批准号: 10078617
• 负责人: MICHAEL A. MARLETTA
• 金额: $ 31.97万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078617
• 关键词: Address; Architecture; Binding; Binding Sites; Biochemical; Biochemistry; Blood Vessels; Cardiovascular Diseases; Cloning; Collaborations; Complement; Complex; Cryoelectron Microscopy; Cyclic GMP; Cysteine; Data; Deuterium; Development; Disease; Electron Microscopy; Enzyme Kinetics; Enzymes; Erectile dysfunction; FDA approved; Functional disorder; Gastrointestinal Diseases; Generations; Global Change; Goals; Grant; Heart Diseases; Heme; Histidine; Homologous Gene; Hydrogen; Hypertension; Iron; Laboratories; Lead; Length; Manduca sexta; Mass Spectrum Analysis; Methods; Modeling; Molecular; Molecular Conformation; Motion; Mutagenesis; Nerve Degeneration; Nitric Oxide; Peptide Mapping; Pharmaceutical Preparations; Pharmacology; Physiological; Planning Techniques; Property; Proteins; Proteomics; Rattus; Roentgen Rays; Signal Transduction; Site; Soluble Guanylate Cyclase; Stroke; Structure; Sulfhydryl Compounds; Vasodilation; Vasodilator Agents; Work; base; chronic thromboembolic pulmonary hypertension; cofactor; drug action; experimental study; heart function; heme a; human disease; in vivo; mutant; neurotransmission; novel therapeutics; pulmonary arterial hypertension; small molecule; synergism; therapeutic target

Nitric oxide (NO) signaling is essential to several physiological functions, and dysfunction in the in this signaling cascade is implicated in multiple diseases such as erectile dysfunction, heart disease, neurodegeneration, stroke, hypertension, and gastrointestinal disease. Soluble guanylate cyclase (sGC) is the primary receptor for NO. NO regulates sGC at two levels and this is consistent with pharmacological observations of NO signaling that are consistent with a two-step activation mechanism by NO. The amplitude and duration of these effects of NO in neuronal signaling, cardiac function, vascular tone and vasodilation are vital to proper function, but the mechanism for the two-step activation by NO has not been thoroughly investigated. A new paradigm for NO signaling through sGC has emerged. Understanding how sGC switches from a low to high activation state is central to this new paradigm. In addition, sGC has become a therapeutic target for the treatment of two forms of pulmonary hypertension: chronic thromboembolic pulmonary hypertension and pulmonary arterial hypertension with the FDA approved Adempas®. Our specific aims include: (i) How does NO activate sGC?, (ii) What is the structural architecture of full-length sGC and what are the inter-domain interactions that contribute to the activation mechanism of sGC?, and (iii) What is the mechanism of action of Adempas® (riociguat) and related stimulators of sGC. Experimental approaches will include the following biochemical methods: enzyme kinetics, cloning, expression, purification and characterization of wild type and site-directed mutants of sGC, electron microscopy structural methods, hydrogen-deuterium exchange and peptide mapping. It is a central goal of this proposal to develop a complete molecular level view of the complex relationship between NO, drugs like Adempas® and sGC. The extension of this work into physiological function will provide a rational basis for the understanding and treatment of NO signaling disorders in human disease.

一氧化氮（NO）信号传导对于多种物理功能至关重要，并且功能障碍 在此信号传导中，级联反应隐含在多种疾病中，例如勃起功能障碍，心脏 疾病，神经变性，中风，高血压和胃肠道疾病。可溶 鸟烯基环化酶（SGC）是NO的主要受体。没有两个级别的SGC调节 这与无信号传导的药物观察一致 两步激活机制NO IN的这些效果的放大器和持续时间 神经元信号传导，心脏功能，血管张力和血管舒张对于正确功能至关重要， 但是，尚未对两步激活的机制进行NO进行彻底研究。一个 已经出现了无信号的新范式。了解SGC如何切换 从低到高激活状态是这种新范式的核心。此外，SGC已成为 治疗两种形式的肺动脉高压的治疗靶标：慢性 血栓栓塞性肺动脉高压和肺动脉高压与FDA 已批准的Adempas®。我们的具体目的包括：（i）如何激活SGC？，（ii）什么是 全长SGC的结构结构以及什么是域间相互作用 有助于SGC的激活机制？和（iii）什么是什么作用机理 SGC的ADEMPAS®（riociguat）和相关刺激剂。实验方法将包括 以下生化方法：酶动力学，克隆，表达，纯化和 SGC，电子显微镜结构的野生型和位置突变体的表征 方法，氢 - 居民交换和肽映射。这是一个核心目标 提出对NO， Adempas®和SGC等药物。将这项工作扩展到生理功能将 为理解和治疗人类的没有信号障碍提供了理性的基础 疾病。