Angiotensin II Receptor Signaling Domains

血管紧张素 II 受体信号传导域

• 批准号: 7536370
• 负责人: Robert WAYNE ALEXANDER
• 金额: $ 34.43万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-18 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7536370
• 关键词: 2-tyrosine; A Mouse; Actins; Adenoviruses; Angiopoietin-2; Angiotensin II; Angiotensin II Receptor; Aorta; Appearance; Binding; Binding Proteins; Biological; Biological Assay; Blood Vessels; Cardiovascular Diseases; Caveolins; Cell Fractionation; Cell membrane; Cells; Chronic; Complex; Confocal Microscopy; Cytoskeleton; Data; Development; Epidermal Growth Factor Receptor; Event; F-Actin; Focal Adhesions; G-Protein-Coupled Receptors; Growth; Growth Factor; HC phosphatase; Hypertension; Hypertrophy; Immigration; Immunofluorescence Immunologic; In Vitro; Infusion procedures; Knockout Mice; Lateral; Life; Light; Location; Mediating; Mediation; Mediator of activation protein; Membrane; Membrane Microdomains; Microtubules; Mitogen-Activated Protein Kinases; Molecular; Movement; Mus; Output; Oxidases; PTPN11 gene; Phosphorylation; Play; Protein Tyrosine Kinase; Proteins; Proto-Oncogenes; Reactive Oxygen Species; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Research Personnel; Role; Signal Transduction; Signaling Molecule; Small Interfering RNA; Smooth Muscle Myocytes; Testing; Transactivation; Transfection; Tyrosine Phosphorylation; catalase; caveolin 1; cellular imaging; human EMS1 protein; in vivo; insight; migration; mutant; new therapeutic target; overexpression; oxidation; programs; protein protein interaction; receptor; response; scaffold; src-Family Kinases; trafficking

Angiotensin II (AngII) is a potent stimulator of vascular hypertrophy which is mediated through the Ang II typel receptor (AT1R) in vascular smooth muscle cells (VSMCs). Many of its important outputs result from transactivation of the EGF receptor (EGF-R), which requires initial activation of cSrc and which is dependent on reactive oxygen species (ROS) derived from NAD(P)H oxidase (Nox). Caveolin-enriched lipid rafts (CE/LRM) are specialized membrane microdomains where signaling molecules such as EGFR are compartmentalized via interacting with caveolin-1 (Cav1). We showed that Ang II promotes AT1R trafficking into CE/LRM, which is required for transactivation of EGF-R and egress of EGF-R from the CE/LRM, resulting in colocalization of pY-EGFR with pY14Cav1 at focal adhesions. We also found that AT1R migration into CE/LRM is dependent on ROS, cSrc, microtubules/actin cytoskeleton and Cav1; however, underlying, organizing molecular mechanisms are poorly understood. cAbl is a F-actin binding non-receptor tyrosine kinase that links receptor tyrosine kinase and actin remodeling. Cav1 is a major ROS-dependent tyrosine phosphorylation target of cAbl. We show that AT1R activates cAbl in VSMCs and mouse aorta, and promotes cAbl binding to the AT1R, which are dependent on ROS, cSrc and Cav1. Preliminary data led us to hypothesize that cAbl functions as a central organizer for ROS, Cav1 and actin cytoskeleton-dependent AT1R trafficking and signaling, which may contribute to vascular hypertrophy. To test this hypothesis, AIM1 will characterize the interaction of AT1R and cAbl, and define the mechanisms of cAbl activation by Ang II. AIM2 will explore the mechanisms by which cAbl is activated by Nox-derived ROS with focusing on oxidative inactivation of SHP-2 which binds directly to AT1R and negatively regulates cSrc. AIMS will explore the molecular mechanisms by which cAbl mediates Ang ll-stimulated AT1R migration into CE/LRM, formation of pY-EGF-R signaling complex at focal adhesions and hypertrophy with focus on cAbl downstream targets, cortactin and Y14Cav1. AIM4 will assess the funcional role of cAbl in Ang ll-induced vascular hypertrophy in vivo. Knockout mice and cells, live cell imaging, cell fractionation, protein-protein interaction and molecular biological aanlysis will be used. These studies should provide new insights into the organization of Ang II signaling and identify potential new targets for novel therapeutic apporaches to cardiovascular diseases.

血管紧张素II（ANGII）是一种有效的血管肥大刺激剂，通过ANG II介导 血管平滑肌细胞（VSMC）中的打字受体（AT1R）。它的许多重要输出来自 EGF受体（EGF-R）的反式激活，该受体需要CSRC的初始激活，而这是依赖性的 在源自NAD（P）H氧化酶（NOX）的活性氧（ROS）上。富含小窝蛋白的脂质筏 （CE/LRM）是特殊的膜微区，其中信号分子（例如EGFR）为 通过与小窝蛋白-1相互作用（CAV1）进行隔离。我们表明，ANG II促进了AT1R贩运 进入CE/LRM，这是EGF-R的EGF-R和EGF-R的EGF-R的反式激活所必需的 导致PY-EGFR与PY14CAV1在焦点粘连下共定位。我们还发现AT1R 迁移到CE/LRM取决于ROS，CSRC，微管/肌动蛋白细胞骨架和CAV1；然而， 基础，组织分子机制的理解很少。 CABL是F-肌动蛋白结合的非受体 酪氨酸激酶将受体酪氨酸激酶和肌动蛋白重塑。 CAV1是主要的ROS依赖性 CABL的酪氨酸磷酸化靶标。我们表明AT1R在VSMC和鼠标主动脉中激活CABL，并且 促进CABL与AT1R的结合，该AT1R取决于ROS，CSRC和CAV1。初步数据导致我们 假设CABL充当ROS，CAV1和肌动蛋白细胞骨架依赖性的中央组织者 AT1R运输和信号传导，可能导致血管肥大。为了检验这一假设，AIM1 将表征AT1R和CABL的相互作用，并定义ANG II的CABL激活的机制。 AIM2将探索由NOX衍生的ROS激活CABL的机制，重点是氧化 SHP-2的失活直接与AT1R结合并负调节CSRC。目标将探索 CABL介导ANG LL刺激的AT1R迁移到CE/LRM的分子机制，形成 PY-EGF-R信号传导复合物在局灶性粘附和肥大上，重点是CABL下游靶标， Cortactin和Y14CAV1。 AIM4将评估CABL在ANG LL诱导的血管肥大中的功能作用 体内。敲除小鼠和细胞，活细胞成像，细胞分馏，蛋白质 - 蛋白质相互作用和分子 将使用生物学溶液。这些研究应为Ang II的组织提供新的见解 信号传导并确定针对心血管疾病的新型治疗性附属物的潜在新靶标。