NO-indepentdent sGC regulation

NO 独立的 sGC 调节

• 批准号: 6945162
• 负责人: NESTOR E FRANCO
• 金额: $ 2.98万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-17 至 2009-08-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6945162
• 关键词: active sites; chemical binding; enzyme activity; enzyme induction /repression; guanylate cyclase; heavy metals; high performance liquid chromatography; mass spectrometry; metals; nitric oxide; oxidation reduction reaction; physiology; predoctoral investigator; thiols

DESCRIPTION (provided by applicant): Although it is clear that NO mediated activation of sGC is the most profound mechanism of regulation, regulation of "basal" or NO-independent sGC activity may be of equal physiological importance. It is the intent of this proposal to investigate other factors regulating sGC such as cellular redox status or metal homeostasis. Our aims are to determine the role of thiol redox chemistry and thlol modification on the activity and regulation of sGC, investigate the role of sGC activation by CO arid NO upon thiol modification and thiol redox chemistry arid to determine the metal binding properties of sGC and the effects of metal binding on activity/regulation. To accomplish our aims we will ascertained the redox status of the cell by measuring both the levels of GSH and the GSH/GSSG ratio. Identical experiments run in parallel will be performed except that cGMP will be measured. The ability of cellular sGC to be activated by NO and CO as a function of cellular redox status will be determined by performing experiments similar to those described above except that NO and CO will be added to the incubations to activate the enzyme. Similarly sGC activity will be determined upon exposed to a variety of metals (e.g. Cu, Zn, Pb, Cd, Ag, As, Hg).

描述（由申请人提供）：虽然很明显NO介导的sGC活化是最深刻的调节机制，但“基础”或NO非依赖性sGC活性的调节可能具有同等的生理重要性。本提案的目的是研究调节sGC的其他因素，如细胞氧化还原状态或金属稳态。我们的目的是确定硫醇氧化还原化学和硫醇修饰对sGC的活性和调节的作用，研究由CO和NO激活的sGC对硫醇修饰和硫醇氧化还原化学的作用，并确定sGC的金属结合性质和金属结合对活性/调节的影响。为了实现我们的目标，我们将通过测量GSH水平和GSH/GSSG比率来确定细胞的氧化还原状态。除了测量cGMP外，将平行进行相同的实验。细胞sGC被NO和CO激活的能力作为细胞氧化还原状态的函数，将通过进行与上述那些相似的实验来确定，除了将NO和CO加入到温育中以激活酶。类似地，将在暴露于各种金属（例如Cu、Zn、Pb、Cd、Ag、As、Hg）后测定sGC活性。