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: 7217328
• 负责人: ANNIE V BEUVE
• 金额: $ 32.44万
• 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 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7217328
• 关键词: Adenylate Cyclase; Allosteric Regulation; Allosteric Site; Amino Acid Sequence; Atherosclerosis; Binding; Binding Sites; Biochemical; Biological; Biology; Cardiovascular system; Catalytic Domain; Cell physiology; Cyclic GMP; Disease; Endothelial Cells; Enzymes; Erectile dysfunction; Functional disorder; Gel Chromatography; Guanosine Triphosphate; Heme; Human; Hypertension; In Vitro; Ion Exchange; Kinetics; Laboratories; Libraries; Measures; Mediating; Modeling; Molecular; Mutation; N-terminal; Nature; Nitric Oxide; Pathway interactions; Peripheral Nervous System; Physiological; Platelet Aggregation Inhibition; Precipitation; Production; Property; Regulation; Research Personnel; Role; Screening procedure; Second Messenger Systems; Signal Pathway; Signal Transduction; Site; Soluble Guanylate Cyclase; Structural Models; Structure; Structure-Activity Relationship; Synaptic plasticity; System; Testing; Thinking; Tissue Extracts; Work; YC-1; analog; base; blood pressure regulation; heme a; in vivo; molecular modeling; mutant; neuropsychiatry; novel; programs; response; second messenger; small molecule libraries

DESCRIPTION (provided by applicant): The cellular processes that are regulated by nitric oxide (NO) are central to many aspects of biology and disease, particularly in the cardiovascular system and the central and peripheral nervous systems. 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 to the heme, the sGC is activated several hundred-fold and it is thought that the NO-cGMP signaling cascade is a key player in regulation of blood pressure, synaptic plasticity and inhibition of platelet aggregation. The proposed studies seek to understand the structural basis of mechanisms of regulation of the sGC and to identify modulators of its activity: 1) Our recent work suggests that sGC contains an allosteric regulatory site. We seek to identify residues that dictate the structure and nature of the proposed allosteric site. Guided by homology with adenylyl cyclases, we conducted a mutational analysis that identifies specific regions of interactions between the two subunits that seem to mediate allosteric activation. We shall characterize these mutants by biochemical studies integrated with molecular modeling. 2) Recent work from our laboratory and others, suggests that there is an endogenous regulator for the sGC. We have discovered but not yet identified a novel endogenous activator. We will identify this activator and screen for others. 3) The wild-type and mutant sGC will be used to test a newly generated chemical library derived from a synthetic activator, YC-1. Active compounds and sGC will be used as templates for molecular modeling. This modeling combined with structure-activity relationship studies will be used to generate detailed and testable hypotheses regarding the physiological modulation of the NO-cGMP signaling pathway. Understanding the mechanisms of regulation of sGC and identifying regulatory molecules will be key to uncovering the molecular basis of some types of hypertension, atherosclerosis and erectile dysfunction.

描述（由申请人提供）：一氧化氮（NO）调节的细胞过程对于生物学和疾病的许多方面都至关重要，特别是在心血管系统以及中枢和周围神经系统中。尽管 NO 的重要性已得到广泛认可，但人们对 NO 受体（可溶性鸟苷酸环化酶 (sGC)）的调节机制知之甚少。 sGC 是一种含血红素的异二聚体，可催化底物 GTP 形成 cGMP。 NO 与血红素结合后，sGC 被激活数百倍，并且人们认为 NO-cGMP 信号级联是调节血压、突触可塑性和抑制血小板聚集的关键因素。拟议的研究旨在了解 sGC 调节机制的结构基础并确定其活性调节剂：1）我们最近的工作表明 sGC 包含一个变构调节位点。我们寻求识别决定所提议的变构位点的结构和性质的残基。在与腺苷酸环化酶同源性的指导下，我们进行了突变分析，确定了似乎介导变构激活的两个亚基之间相互作用的特定区域。我们将通过生化研究与分子建模相结合来表征这些突变体。 2) 我们实验室和其他实验室最近的工作表明，sGC 存在内源性调节因子。我们已经发现但尚未鉴定出一种新型内源性激活剂。我们将为其他人识别该激活器并进行筛选。 3) 野生型和突变型 sGC 将用于测试源自合成激活剂 YC-1 的新生成的化学文库。活性化合物和 sGC 将用作分子建模的模板。该模型与结构-活性关系研究相结合，将用于生成有关 NO-cGMP 信号通路生理调节的详细且可测试的假设。了解 sGC 的调节机制并识别调节分子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。