Regulation of guanylyl cyclase

鸟苷酸环化酶的调节

• 批准号: 6640327
• 负责人: FERID MURAD
• 金额: $ 40.11万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6640327
• 关键词: apoptosis; biological signal transduction; cell differentiation; cyclic GMP; cytotoxicity; enzyme activity; enzyme linked immunosorbent assay; enzyme mechanism; gel mobility shift assay; genetic transcription; guanylate cyclase; intracellular; messenger RNA; neuroblastoma; nitric oxide; oxidative stress; polymerase chain reaction; second messengers; site directed mutagenesis; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Soluble guanylyl cyclase (sGC) is one of the crucial enzymes in the NO-cyclic GMP (cGMP) signaling cascade involved in smooth muscle relaxation, inhibition of platelet aggregation, and neurotransmission. sGC is the target of various pharmacological compounds currently used for treatment of cardiovascular disorders. However, the mechanisms governing stimulation of activity or expression of sGC are poorly understood. Effective manipulation of sGC-dependent synthesis of cGMP for therapeutic purposes requires understanding of the pathways involved in regulation of sGC levels and activity. The goal of this proposal is to determine the molecular mechanism of enzyme activation by alisoteric effectors including development of approaches for NO-independent regulation of sGC, investigation of the mechanisms regulating sGC expression, and analysis of the effects of short-term and long-term modulation of sGC activity and expression on cell physiology. To achieve this goal, a constitutively activated sGC mutant will be expressed in a cell model providing for elevated levels of cGMP, whereas expression of domains involved in formation of functional heterodimers of sGC will result in decreased enzyme levels and lowered NO-dependent synthesis of cGMP. The promoter fragments of sGC genes will be identified and characterized as well as the mechanisms of post-transcriptional regulation of sGC Human cell culture models will be developed and the effects of short-term and long-term sGC regulation on cell physiology will be determined. The knowledge of the mechanism of sGC regulation will advance our understanding of the role of sGC/cGMP system in various biological processes.

描述（由申请人提供）：可溶性鸟苷酸环化酶（sGC）是涉及平滑肌松弛、血小板聚集抑制和神经传递的NO-环鸟苷酸（cGMP）信号级联中的关键酶之一。 sGC是目前用于治疗心血管疾病的多种药理化合物的靶点。然而，人们对刺激 sGC 活性或表达的机制知之甚少。出于治疗目的，有效控制 sGC 依赖性 cGMP 合成需要了解参与 sGC 水平和活性调节的途径。本提案的目标是确定异位效应子激活酶的分子机制，包括开发不依赖于 NO 的 sGC 调节方法、研究调节 sGC 表达的机制，以及分析短期和长期调节 sGC 活性和表达对细胞生理学的影响。为了实现这一目标，组成型激活的 sGC 突变体将在细胞模型中表达，从而提供升高的 cGMP 水平，而参与 sGC 功能性异二聚体形成的结构域的表达将导致酶水平降低并降低 NO 依赖性 cGMP 合成。将鉴定和表征 sGC 基因的启动子片段，以及 sGC 转录后调控的机制。将开发人类细胞培养模型，并确定短期和长期 sGC 调控对细胞生理学的影响。对sGC调节机制的了解将有助于我们更好地理解sGC/cGMP系统在各种生物过程中的作用。