BRADYKININ SIGNALING IN REGULATION OF eNOS

eNOS 调节中的缓激肽信号传导

• 批准号: 6594510
• 负责人: RICHARD C VENEMA
• 金额: $ 31.13万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-15 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6594510
• 关键词: SDS polyacrylamide gel electrophoresis; arginine; biological signal transduction; blood pressure; bradykinin; caveolins; confocal scanning microscopy; enzyme activity; heat shock proteins; immunoprecipitation; laboratory mouse; nitric oxide; nitric oxide synthase; oxidation; phosphorylation; polymerase chain reaction; protein kinase; protein protein interaction; tissue /cell culture; vascular endothelium; vasodilation

DESCRIPTION (provided by applicant): Endothelium-derived nitric oxide (NO) has a crucial role in regulation of the state of vasodilation of blood vessels and hence in regulation of blood pressure. Furthermore, because NO regulates this and a number of other important vascular processes, abnormalities in vascular NO production are thought to contribute to the pathogenesis of certain vascular disorders such as those of atherosclerosis, diabetes, and hypertension. NO is synthesized in endothelial cells by oxidation of L-arginine in a reaction catalyzed by the enzyme, endothelial nitric oxide synthase (eNOS). Recent investigations in several laboratories have established a role for Ser-1179 phosphorylation and Thr-497 dephosphorylation in agonist regulation of eNOS activity in endothelial cells. Data presented in the Preliminary Studies section of this proposal provide evidence for two additional sites of eNOS phosphorylation at Ser-617 and Ser-635. Our preliminary data shows that these two sites are transiently phosphorylated in cultured endothelial cells in response to stimulation with the eNOS-activating agonists bradykinin (BK), ATP, and vascular endothelial growth factor. The principal aim of the present proposal is to examine the hypothesis that eNOS activity in vascular endothelial cells is regulated in part by BK-stimulated phosphorylation of the enzyme at Ser-617 and Ser-635. Additional aims are to elucidate the molecular mechanism(s) by which phosphorylation at Ser-617 and Ser-635 alters enzyme catalytic activity and to test the hypothesis that phosphorylation at Ser-617 or Ser-635 alters eNOS protein-protein interactions with caveolin-1, the bradykinin B2 receptor, or heat shock protein 90. Finally, we will examine whether certain pathophysiological conditions, such as hyperglycemia or oxidative stress, alter either basal or BK-stimulated phosphorylation of eNOS at Ser-617 or Ser-635.

描述（由申请人提供）：内皮衍生的一氧化氮（NO）在调节血管血管血管血管状态的状态中至关重要，因此在调节血压方面具有至关重要的作用。此外，由于没有调节这一点和许多其他重要的血管过程，因此血管异常无产生被认为有助于某些血管疾病的发病机理，例如动脉粥样硬化，糖尿病和高血压。通过在酶，内皮一氧化氮合酶（ENOS）催化的反应中氧化L-精氨酸在内皮细胞中合成的NO。最近对几个实验室的研究已经确立了Ser-1179磷酸化和THR-497去磷酸化在内皮细胞中eNOS活性的激动剂调节中的作用。本提案的初步研究部分中提供的数据为SER-617和SER-635的ENOS磷酸化其他两个地点提供了证据。我们的初步数据表明，这两个位点在培养的内皮细胞中瞬时磷酸化，以响应于eNOS激活的激动剂刺激性激动剂（BK），ATP和血管内皮生长因子。 本提案的主要目的是检查以下假设：血管内皮细胞中的eNOS活性部分通过Ser-617和Ser-635的BK刺激酶的磷酸化的部分调节。 Additional aims are to elucidate the molecular mechanism(s) by which phosphorylation at Ser-617 and Ser-635 alters enzyme catalytic activity and to test the hypothesis that phosphorylation at Ser-617 or Ser-635 alters eNOS protein-protein interactions with caveolin-1, the bradykinin B2 receptor, or heat shock protein 90. Finally, we will examine whether certain病理生理状况，例如高血糖或氧化应激，在Ser-617或Ser-635上改变了基底或BK刺激的eNOS磷酸化。