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) 信号传导对于多种生理功能和功能障碍至关重要 这种信号级联与多种疾病有关，例如勃起功能障碍、心脏疾病 疾病、神经退行性疾病、中风、高血压和胃肠道疾病。 鸟苷酸环化酶 (sGC) 是 NO 的主要受体，在两个水平上调节 sGC 这与 NO 信号传导的药理学观察结果一致，NO 信号传导与 NO 的两步激活机制 NO 的这些作用的幅度和持续时间。 神经信号、心脏功能、血管张力和血管舒张对于正常功能至关重要， 但NO的两步激活机制尚未得到彻底研究。 通过 sGC 进行 NO 信号传导的新范例已经出现，了解 sGC 如何切换。 从低激活状态到高激活状态是这个新范例的核心。此外，sGC 已成为这一新范式的核心。 治疗两种形式肺动脉高压的治疗目标：慢性 FDA 治疗血栓栓塞性肺动脉高压和肺动脉高压 批准的 Adempas® 我们的具体目标包括：(i) NO 如何激活 sGC？，(ii) 什么是 全长 sGC 的结构体系以及域间相互作用是什么 有助于 sGC 的激活机制？ (iii) 的作用机制是什么 Adempas® (riociguat) 和相关 sGC 刺激剂将包括 以下生化方法：酶动力学、克隆、表达、纯化和 sGC野生型和定点突变体的表征，电子显微镜结构 方法、氢-氘交换和肽谱分析是其核心目标。 建议开发 NO 之间复杂关系的完整分子水平视图， 像 Adempas® 药物和 sGC 一样，这项工作将扩展到生理功能。 为人类NO信号传导障碍的理解和治疗提供合理依据 疾病。