Local Signaling in Diabetic Comorbidities

糖尿病合并症的局部信号传导

• 批准号: 8891765
• 负责人: John D Scott
• 金额: $ 33.96万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-10 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8891765
• 关键词: A kinase anchoring protein; Adult; Adverse effects; Animals; Arteries; Binding Sites; Blood Pressure; Blood Vessels; Calcineurin; Calcium; Cells; Child; Comorbidity; Complex; Complications of Diabetes Mellitus; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Diabetes Mellitus; Diffuse; Electron Microscopy; Enzymes; Event; Exhibits; Eye diseases; Family; Functional disorder; Goals; Higher Order Chromatin Structure; Hormones; Human; Hypertension; Image; Individual; Insulin; Investigation; Ion Channel; Kidney Failure; L-Type Calcium Channels; Location; Measures; Mediating; Metabolic; Metabolic syndrome; Molecular; Multienzyme Complexes; Mus; Muscle Cells; Orthologous Gene; Pancreas; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Photobleaching; Physiological; Population; Protein Isoforms; Protein Kinase; Protein Kinase C; Protein phosphatase; Proteins; Regulation; Regulatory Element; Research; Resolution; Risk; Role; Shapes; Signal Transduction; Signal Transduction Pathway; Smooth Muscle; Stretching; Stroke; Testing; Tissues; United States; Vascular Smooth Muscle; Work; blood glucose regulation; carbohydrate metabolism; combat; constriction; diabetic; diabetic cardiomyopathy; lipid metabolism; macromolecular assembly; mouse model; particle; programs; protein complex; protein protein interaction; public health relevance; single molecule; vasoconstriction

 DESCRIPTION (provided by applicant): The spatial and temporal organization of signal transduction pathways influences the precision and fidelity of intracellular events. We have discovered a family of non-catalytic regulatory elements called A-Kinase Anchoring Proteins (AKAPs) that bring together different combinations of calcium- and cAMP-responsive protein kinases and phosphatases to customize the regulation of effector proteins. This application focuses on the molecular pathophysiology of a multivalent anchoring protein known as AKAP79/150 (AKAP79 is the human form, AKAP150 is the murine ortholog). We discovered this protein and defined binding sites for protein kinase A (PKA), the phosphatase PP2B and conventional isoforms of protein kinase C (PKC). Physiological studies have demonstrated that AKAP79/150-tethered enzymes actively participate in the control of glucose homeostasis and coordinate certain extra pancreatic aspects of insulin action. This application is developed around exciting data pertaining to the role of anchored PKC in signaling events that govern arterial constriction and hypertension. Molecular events underlying this pathological state include aberrant calcium influx through ion channels that are controlled, in part, by AKAP79/150-associated PKC. The hypothesis to be tested is that manipulation of anchored PKC affords a measure of vascular benefit to alleviate diabetes-induced hypertension. Two specific aims are proposed: Aim 1: How are individual AKAP79 complexes configured? We will harness three cutting-edge approaches: a) single-molecule pull-down photobleaching (SiMPull) to calculate the range of enzyme combinations on individual anchoring proteins, b) single-particle electron microscopy (EM) to obtain near-atomic structures of higher-order AKAP79 assemblies and c) super resolution imaging of native AKAP-enzyme complexes in vascular smooth muscle. Aim 2: Does PKC-anchoring govern diabetes-related hypertension? Sixty-five percent of diabetics develop hypertension. Our AKAP150-/- and AKAP150ΔPKC knockin mice exhibit lower basal blood pressure suggesting that AKAP-associated PKC participates in the control of vascular tone. Aim 2 will test if selective disruption of this protein-protein interactin reduces vascular tone in mouse models of diabetic hypertension.

 描述（由申请人提供）：信号转导途径的空间和时间组织影响细胞内事件的精确度和保真度。我们已经发现了一个称为A-激酶调节蛋白（AKAP）的非催化调节元件家族，它们将钙和cAMP响应性蛋白激酶和磷酸酶的不同组合结合在一起，以定制效应蛋白的调节。本申请集中于称为AKAP 79/150的多价锚定蛋白的分子病理生理学（AKAP 79是人形式，AKAP 150是鼠直系同源物）。我们发现了这种蛋白质，并确定了蛋白激酶A（PKA），磷酸酶PP 2B和蛋白激酶C（PKC）的常规亚型的结合位点。生理学研究表明，AKAP 79/150-栓系酶积极参与葡萄糖稳态的控制，并协调胰岛素作用的某些胰腺外方面。这个应用程序是围绕着激动人心的数据开发的锚定蛋白激酶C在信号事件中的作用，支配动脉收缩和高血压。这种病理状态下的分子事件包括通过离子通道的异常钙内流，其部分由AKAP 79/150相关的PKC控制。有待检验的假设是，操纵锚定的PKC提供了一个衡量血管的好处，以减轻糖尿病引起的高血压。提出了两个具体的目标：目标1：如何个别AKAP 79复合物配置？我们将利用三种尖端方法：a）单分子下拉光漂白（SiMPull）计算单个锚定蛋白上酶组合的范围，B）单粒子电子显微镜（EM）获得高阶AKAP 79组装的近原子结构和c）血管平滑肌中天然AKAP-酶复合物的超分辨率成像。目的2：PKC锚定是否控制糖尿病相关高血压？65%的糖尿病患者会发展为高血压。我们的AKAP 150-/-和AKAP 150 ΔPKC敲入小鼠表现出较低的基础血压，表明AKAP相关的PKC参与了血管张力的控制。目标2将测试在糖尿病高血压小鼠模型中选择性破坏这种蛋白质-蛋白质相互作用是否会降低血管张力。