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是核心，尤其是在心血管系统以及中心和周围神经系统中。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是一种含血红素的异二聚体，可催化底物GTP的CGMP形成。 NO与血红素结合后，SGC被激活了几百倍，并且认为No-CGMP信号级联反应是调节血压，突触可塑性和抑制血小板聚集的关键参与者。拟议的研究试图了解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包含一个变构调节部位。我们寻求确定决定拟议变构位点的结构和性质的残留物。在与腺苷循环酶同源性的指导下，我们进行了突变分析，该突变分析确定了两个亚基之间的相互作用特定区域，这些区域似乎介导了变构激活。我们将通过与分子建模集成的生化研究来表征这些突变体。 2）我们实验室和其他实验室的最新工作表明，SGC有一个内源性调节剂。我们已经发现但尚未确定一种新型的内源性激活剂。我们将确定此激活剂并为他人屏幕。 3）野生型和突变体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。这种建模与结构 - 活性关系研究的结合将用于生成有关NOCGMP信号通路的生理调节的详细和可检验的假设。了解SGC调节和识别调节分子的机制将是发现某些类型的高血压，动脉粥样硬化和勃起功能障碍的分子基础的关键。