INSULIN RECEPTOR SUBSTRATE -1 AND FLUID SHEAR STRESS

胰岛素受体底物 -1 和流体剪切应力

• 批准号: 6536643
• 负责人: Francis Kim
• 金额: $ 12.5万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6536643
• 关键词: SDS polyacrylamide gel electrophoresis; biological signal transduction; enzyme activity; fluid flow; high performance liquid chromatography; human genetic material tag; human tissue; immunoprecipitation; insulin receptor; mass spectrometry; molecular cloning; nitric oxide; phosphorylation; physiologic stressor; protein binding; protein kinase; receptor binding; shear stress; site directed mutagenesis; thin layer chromatography; tissue /cell culture; tumor necrosis factor alpha; vascular endothelium; western blottings

(Adapted from applicant's abstract) The goals of the proposed research project are: 1) to prepare and train the candidate for a career in cardiovascular research by providing him with the necessary education, environment, and support, and to 2) to gain insight into the potential role of IRS-1 (insulin receptor substrate) in signal transduction of shear stress induced stimulation of NO production and the modulation of IRS-1 signaling by TNF-alpha. The candidate will be provided with extensive education in cardiovascular biochemistry, signal transduction, biophysics, research methods, and statistics. The candidate will be supported and mentored by Marshall Corson and Ruedi Aebersold who with the research advisory committee will monitor the candidate's progress. Finally, the Department of Medicine and Division of Cardiology will provide full commitment and support to developing the candidate's career as an independent investigator. The proposed research project is the foundation of attaining these goals. IRS-1 is a multi domain cytosolic protein that plays an important role in insulin signal transmission and has recently been shown to be used by multiple receptors for cytokines, growth hormones, interleukin and inteferons. We have obtained preliminary data implicating IRS-1 in mechanotranduction of FSS (fluid shear stress) in endothelial cell NO production. The major goal of the research proposal is to determine the molecular mechanisms regulating IRS-1 in this context. Type II diabetes is prevalent in the United States, affecting over 5 percent of the population. Type II diabetes involves defects in insulin action as well as insulin secretion. Insulin resistance is a state in which target cells fail to respond to ordinary levels of circulating insulin; at the molecular level, modifications in the insulin receptor (IR) or IRS-1 can lead to insulin resistance. TNF-alpha, which increases the serine phosphorylation of IRS-1, diminishes insulin induced tyrosine phosphorylation of IRS-1 and this modification of IRS-1 has recently been proposed as an important mediator of insulin resistance. Endothelial cells are not known to play a significant role in glucose metabolism and the role of IRS-1 in FSS endothelial NO signaling has not been previously studied. Our laboratory has recently reported that mechanotransduction of FSS in No production utilizes a P13-Kinase dependent pathway to phosphorylate eNOS. We hypothesize that shear stress induced NO production and insulin signaling share overlapping elements of the same signaling pathway and the same docking protein IRS-1 and that both pathways are modulated by TNF-alpha which increases the serine phosphorylation of IRS- 1. To further characterize this potential link between insulin and NO signaling. Specific Aim 3: Characterize FSS dependent signaling elements which associate with IRS-1.

（改编自申请人摘要）拟议研究项目的目标 是：1）准备和培训候选人的职业生涯在心血管 通过为他提供必要的教育、环境和 支持，以及2）深入了解IRS-1（胰岛素）的潜在作用 受体底物）在切应力刺激信号转导中的作用 NO的产生和TNF-α对IRS-1信号的调节。 的 候选人将获得广泛的心血管教育 生物化学，信号转导，生物物理学，研究方法， 统计 候选人将得到马歇尔科森的支持和指导 和Ruedi Aebersold谁与研究咨询委员会将监测 候选人的进步。 最后，医学系和 心脏病学将提供全面的承诺和支持， 候选人作为独立调查员的职业生涯。 拟议研究 项目是实现这些目标的基础。 IRS-1是一种多结构域的胞质蛋白，其在细胞凋亡中起重要作用。 胰岛素信号传输，并且最近已经显示出被多个 细胞因子、生长激素、白细胞介素和干扰素的受体。 我们有 获得的初步数据暗示IRS-1在FSS的机械转导 （流体剪切应力）在内皮细胞NO产生中的作用。 的主要目标 研究计划是确定IRS-1的分子调控机制， 这个背景。 II型糖尿病在美国很普遍， 的人口。 II型糖尿病涉及胰岛素作用缺陷， 以及胰岛素分泌。 胰岛素抵抗是一种状态， 细胞不能对正常水平的循环胰岛素作出反应; 在分子水平上，胰岛素受体（IR）或IRS-1的修饰可以导致 胰岛素抵抗 TNF-α，它增加丝氨酸磷酸化， 降低胰岛素诱导的IRS-1酪氨酸磷酸化， IRS-1的这种修饰最近被认为是一种重要的媒介物， 胰岛素抵抗。 内皮细胞在葡萄糖代谢中起重要作用 IRS-1在FSS内皮NO信号传导中的作用尚未被证实。 以前研究过。 我们的实验室最近报告说， 在NO产生中，FSS的机械转导利用P13激酶依赖的 磷酸化eNOS的途径。 我们假设剪切应力诱导NO 产生和胰岛素信号传导共享相同的重叠元件， 信号通路和相同的对接蛋白IRS-1， 受TNF-α调节，从而增加IRS的丝氨酸磷酸化- 1. 为了进一步描述胰岛素和NO之间的潜在联系， 发信号。 具体目标3：表征FSS依赖性信号传导元件 与IRS-1相关的