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; 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（通过应用程序提供）：信号转导途径的空间和临时组织会影响细胞内事件的精确性和保真度。我们发现了一个称为A-激酶锚定蛋白（AKAP）的非催化调节元件的家族，该元件将钙和cAMP反应性蛋白激酶和磷酸酶的不同组合汇集在一起​​，以自定义效应蛋白的调节。该应用集中在多价锚定蛋白AKAP79/150的分子病理生理上（AKAP79是人类形式，AKAP150是鼠类直系同源物）。我们发现了蛋白激酶A（PKA），磷酸酶PP2B和蛋白激酶C（PKC）的常规同工型的这种蛋白质和定义的结合位点。生理研究表明，AKAP79/150螺旋酶积极参与葡萄糖稳态和坐标的控制。胰岛素作用的某些额外胰腺方面。该应用是围绕与锚定PKC在控制伪影和高血压的信号事件中的作用有关的令人兴奋的数据开发的。这种病理状态的基础事件包括通过AKAP79/150相关PKC控制的离子通道的异常钙影响。要测试的假设是，对锚定PKC的操纵为减轻糖尿病引起的高血压的血管益处提供了衡量。提出了两个具体目标：目标1：如何配置单个AKAP79复合物？ 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：PKC锚定是否控制着与糖尿病相关的高血压？百分之六十五的糖尿病患者发展高血压。我们的AKAP150 - / - 和AKAP150ΔPKC敲击蛋白暴露于较低的碱性血压，这表明与AKAP相关的PKC参与了血管张力的控制。 AIM 2将测试该蛋白质蛋白相互作用蛋白的选择性破坏是否会降低糖尿病高血压小鼠模型中的血管张力。