Nitric Oxide Signaling And Soluble Guanylate Cyclase

一氧化氮信号传导和可溶性鸟苷酸环化酶

• 批准号: 7728873
• 负责人: MICHAEL A. MARLETTA
• 金额: $ 29.74万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7728873
• 关键词: Acute; Allosteric Site; Binding Sites; Biochemical; Biochemistry; Blood Vessels; Cardiac; Cardiovascular Physiology; Cell model; Complex; Disease; Doctor of Philosophy; Electrons; Enzyme Activation; Enzymes; Erectile dysfunction; Functional disorder; Gastrointestinal Diseases; Goals; Guanosine Triphosphate; Heart Diseases; Heme; Hypertension; Kinetics; Maps; Mass Spectrum Analysis; Methods; Minor; Molecular; Nature; Nerve Degeneration; Neuraxis; Neurons; Nitric Oxide; Nucleotides; Paper; Pharmacology; Physiological; Postdoctoral Fellow; Published Comment; Reaction; Regulation; Research Personnel; Role; Signal Transduction; Site; Soluble Guanylate Cyclase; Stroke; Substrate Specificity; System; Time; Ursidae Family; Vasodilation; Work; base; expression cloning; graduate student; human disease; in vivo; mutant; novel therapeutics; oxidative damage; protein crosslink; receptor; research study

Nitric oxide signaling is critical 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. Activation and deactivation of soluble guanylate cyclase (sGC) is of central importance in nitric oxide (NO) signaling. NO regulates sGC at two levels and this is consistent with numerous pharmacological observations of NO signaling that describe tonic and acute roles for NO. The amplitude and duration of these effects of NO in neuronal signaling, cardiac function, vascular tone and vasodilation are vital to the proper function of these systems, but the mechanism for two NO effects 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. Our specific aims include: (i) Characterization of NO activation of sGC, with emphasis on studies of the physiological relevance of low and high activity states, (ii) Characterization of the allosteric nucleotide and activator binding site(s), and the role of nucleotide in modulating NO activation of the enzyme, and (iii) determining the effect of oxidative damage to sGC and the role of this in human disease. Experimental approaches will include physical biochemical methods such as mass spectrometry and rapid-reaction kinetics, cloning, expression, purification and characterization of wild type and sitedirected mutants of sGC, and experiments in various cellular systems to extend the findings into an in vivo setting. It is a central goal of this proposal to develop an entirely new understanding of the complex relationship between NO and sGC. We seek to develop a complete molecular level view of sGC activation and deactivation by NO and nucleotides (ATP and GTP). 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， 心脏病、神经退行性疾病、中风、高血压和胃肠道疾病。 可溶性鸟苷酸环化酶 (sGC) 的激活和失活至关重要 一氧化氮（NO）信号传导。 NO 在两个水平上调节 sGC，这与 描述强直性和急性性的 NO 信号传导的大量药理学观察 NO. 的角色NO 对神经元信号传导的影响的幅度和持续时间， 心脏功能、血管张力和血管舒张对于这些功能的正常发挥至关重要 系统，但两种 NO 效应的机制尚未得到彻底研究。一个 通过 sGC 进行 NO 信号传导的新范例已经出现。了解 sGC 的原理 从低激活状态到高激活状态的切换是这种新范例的核心。我们的具体 目标包括： (i) 表征 sGC 的 NO 激活，重点研究 低和高活动状态的生理相关性，(ii) 的表征 变构核苷酸和激活剂结合位点，以及核苷酸在 调节酶的 NO 活化，以及 (iii) 确定氧化作用的影响 sGC 的损伤及其在人类疾病中的作用。实验方法将 包括物理生化方法，例如质谱法和快速反应 野生型和定点的动力学、克隆、表达、纯化和表征 sGC 的突变体，并在各种细胞系统中进行实验以扩展 体内环境中的发现。该提案的核心目标是开发一个完整的 对 NO 和 sGC 之间复杂关系的新认识。我们力求 开发 NO 和 sGC 激活和失活的完整分子水平视图 核苷酸（ATP 和 GTP）。这项工作扩展到生理功能将 为理解和治疗NO信号紊乱提供合理基础 人类疾病。