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（由申请方提供）：可溶性鸟苷酸环化酶（sGC）是参与平滑肌松弛、血小板聚集抑制和神经传递的NO-环GMP（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各种药理学化合物的靶标。然而，调控sGC活性或表达的刺激的机制知之甚少。为了治疗目的而对sGC依赖性cGMP合成的有效操纵需要理解参与调节sGC水平和活性的途径。该提案的目标是确定酶激活的分子机制，包括开发的方法NO-非依赖性的调节sGC，调查的机制调节sGC的表达，并分析短期和长期的调节sGC的活性和表达对细胞生理学的影响。为了实现该目标，组成型激活的sGC突变体将在提供升高的cGMP水平的细胞模型中表达，而参与sGC的功能性异二聚体形成的结构域的表达将导致酶水平降低和cGMP的NO依赖性合成降低。将鉴定和表征sGC基因的启动子片段以及sGC的转录后调控机制。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调控机制的了解将有助于我们进一步理解sGC/cGMP系统在各种生物过程中的作用。