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p120 Catenin and Endothelial Monolayer Function

p120 连环蛋白和内皮单层功能

基本信息

批准号：
7761679
负责人：
PETER A VINCENT
金额：
$ 34.52万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-02-01 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7761679
关键词：
Actins Acute Lung Injury Adherens Junction Atherosclerosis Binding Blood Vessels Cadherins Cell Adhesion Molecules Cell Fractionation Cell Nucleus Cell physiology Cell-Cell Adhesion Cells Complex Cytoplasm Cytoplasmic Tail Cytoskeleton Diabetes Mellitus Disease Doctor of Philosophy Edema Endothelial Cells Etiology Extracellular Domain Figs - dietary Functional disorder Growth Factor Guanosine Triphosphate Phosphohydrolases Inflammation Mediators Inflammatory Intervention Introns Lead Membrane Molecular Monomeric GTP-Binding Proteins Myocardial Infarction N-Cadherin Pathway interactions Phenotype Phosphorylation Phosphotransferases Play Post-Translational Modification Site Process Proteins Publications Pulmonary Edema Recombinant Proteins Recombinants Regulation Research Role Scaffolding Protein Serine Signal Pathway Signal Transduction Signaling Molecule Site Small Interfering RNA Stroke Structure Threonine Transcriptional Regulation VAV2 gene armadillo proteins cadherin 5 insight member monolayer mutant numb protein scaffold src-Family Kinases

项目摘要

Dysfunction of the endothelial cell barrier has been implicated in the etiology of a variety of disease states including atherosclerosis, microvascular hyperpermeability with diabetes and pulmonary edema following acute lung injury. VE-cadherin is a cell-cell adhesion protein that participates in the regulation of a number of endothelial cell functions. The cytoplasmic domain of VE-cadherin binds to the armadillo family of proteins called catenins to form the adherens junction (AJ). Changes in the phosphorylation of AJ proteins have been implicated in regulating barrier function. For example, phosphorylation of AJ proteins occurs during the formation of edema following stroke and myocardial infarction, a process that is dependent on activation of Src kinase. This proposalfocuses on p120, a catenin that binds to the juxtamembrane of VE- cadherin where it serves a scaffolding role for a number of proteins. Interestingly, p120 is a target for Src kinase and changes in the phosphorylation state of p120 are associated with decreased endothelial barrier function following treatment with inflammatory mediators. In addition, our recent publications have shown that the interaction of p120 with VE-cadherin is required for maintaining VE-cadherin levels in endothelial cells. Within this context, we hypothesize that the juxtamembrane region of VE-cadherin and its interaction with p120 plays a critical role in regulating cell-cell adhesion and barrier function in endothelial cell monolayers. This hypothesis will be investigated by performing three specific aims: 1) identify the regions of VE-cadherin that are responsible for maintaining VE-cadherin levels and barrier function in endothelial cell monolayers; 2) determine if p120 serves as a signaling molecule when released from the membrane into the cytoplasm; and 3) identify the domains of p120 required for the regulation of endothelial cell-cell adhesion and barrier function. We will use expression of chimeric molecules of VE and N-cadherin as well as deletion mutants of p120 to perform structure/function analysis of p120 and VE-cadherin and their effects on endothelial cell-cell adhesion and barrier function. Expression of these molecules will be performed in the absence of endogenous cadherin or p120 that will be depleted using siRNA. These studies will provide new insights into the cellular and molecular mechanisms controlling endothelial barrier function and potentially identify new sites for intervention.

内皮细胞屏障功能障碍与多种疾病的病因有关包括动脉粥样硬化、糖尿病微血管高通透性和随后的肺水肿急性肺损伤。Ve-钙粘蛋白是一种细胞-细胞黏附蛋白，参与细胞数量的调节。内皮细胞的功能。VE-钙粘附素的胞质结构域与称为连环蛋白的蛋白质形成粘着连接(AJ)。AJ蛋白磷酸化的变化与调节屏障功能有关。例如，AJ蛋白发生了磷酸化。在中风和心肌梗塞后形成水肿的过程中，这一过程依赖于 Src激酶的激活。这一建议关注的是p120，一种与VE膜旁结合的连锁素- 钙粘附素是一种为许多蛋白质提供支架的物质。有趣的是，p120是Src的靶点蛋白激酶和p120磷酸化状态的改变与内皮屏障降低有关炎症介质治疗后的功能。此外，我们最近的出版物显示， P120与VE-钙粘蛋白的相互作用是维持内皮细胞VE-钙粘蛋白水平所必需的细胞。在此背景下，我们假设VE-钙粘蛋白的膜旁区域及其相互作用 P120在调节内皮细胞的细胞间黏附和屏障功能中起关键作用单层。这一假设将通过执行三个具体目标来进行研究：1)确定区域在内皮细胞中负责维持VE-钙粘蛋白水平和屏障功能的VE-钙粘蛋白的表达细胞单层；2)确定当p120从膜上释放时是否作为信号分子进入细胞质；以及3)确定内皮细胞-细胞调节所需的p120结构域具有粘合和阻隔功能。我们还将利用VE和N-钙粘蛋白嵌合分子的表达作为p120的缺失突变体进行p120和VE-钙粘附素的结构/功能分析及其影响对内皮细胞-细胞黏附和屏障功能的影响。这些分子的表达将在缺乏内源性钙粘附素或p120，它们将通过siRNA被耗尽。这些研究将提供对控制内皮屏障功能的细胞和分子机制的新见解有可能确定新的干预地点。