Coupling of vascular CaV1.2 channels in health and disease

血管 CaV1.2 通道在健康和疾病中的耦合

• 批准号: 10451644
• 负责人: Manuel F Navedo
• 金额: $ 57.29万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10451644
• 关键词: Active Biological Transport; Acute; Address; Adrenergic Agents; Amino Acids; Animal Model; Animals; Arteries; Biochemistry; Blood Pressure; Blood Vessels; Blood flow; C-terminal; Calcineurin; Cell Nucleus; Cell physiology; Cells; Chemosensitization; Chronic; Complex; Coupled; Coupling; Cyclic AMP-Dependent Protein Kinases; Data; Development; Diabetes Mellitus; Diffusion; Disease; Electrophysiology (science); Exposure to; Fostering; Foundations; Gene Expression; Glucose; Goals; Health; Human; Ion Channel; Knowledge; Lasers; Lateral; Mediating; Membrane; Membrane Proteins; Microscopy; Mission; Modeling; Molecular; Mus; Muscle Cells; Myography; Nanoscopy; Neurons; Optics; Pathology; Phosphorylation; Phosphorylation Site; Physiological; Physiology; Play; Process; Proteins; Public Health; Regulation; Risk Factors; Role; Signal Pathway; Signal Transduction; Site; Smooth Muscle Myocytes; Source; Surface; Techniques; Testing; Therapeutic; United States National Institutes of Health; Vesicle; Work; base; diabetic patient; experimental study; extracellular; health goals; in silico; innovation; nano; nanoscale; non-diabetic; novel; novel therapeutic intervention; particle; patch clamp; pressure; response; tool; trafficking; transcription factor NF-AT c3; treatment strategy; vasoconstriction; voltage

Abstract . How a given protein organizes into a functional complex in health and disease is particularly relevant for membrane proteins that serve as key information entry points for cells. A suitable and highly relevant example is voltage-gated L-type CaV1.2 channels, which play a major role in arterial myocyte function and vascular reactivity. These channels have been shown to gate in unison (i.e. cooperative gating) to amplify Ca2+ influx. At present, however, a comprehensive understanding of mechanisms fostering the induction of CaV1.2 cooperative gating as well as its functional implications in health and disease represent major knowledge gaps. The overall objective of this proposal is to investigate the requirement and physiological consequences whereby phosphorylation of a single amino acid – S1928 – in the C-terminal of vascular CaV1.2 channels promotes dynamic spatial organization of CaV1.2 to facilitate cooperative gating at the surface membrane. To accomplish this goal, we are testing the central hypothesis that CaV1.2 S1928 phosphorylation tunes dynamic channel clustering and cooperative gating, and that this contributes to modulate vascular function in response to elevated extracellular glucose and during diabetes. This hypothesis is formulated on the basis of strong and rigorous preliminary data revealing an unanticipated and remarkable role for S1928 phosphorylation as the culprit for redistribution and assembly of CaV1.2 subunits into superclusters at the surface membrane of arterial myocytes upon elevated glucose and diabetes. CaV1.2 superclusters mediated by S1928 phosphorylation promotes CaV1.2 cooperative gating and Ca2+ influx amplification into arterial myocytes. Key findings that further underscore the significance of our observations is that CaV1.2 S1928 phosphorylation is necessary for activation of prohypertensive signaling pathways, vasoconstriction and altered blood flow upon elevated glucose and during diabetes. Moreover, critical observations have been validated in freshly dissociated human arterial myocytes from nondiabetic and diabetic patients, underscoring the translational relevance. Emerging and innovative concepts that will be explored in this application are the role of S1928 phosphorylation as 1) a rheostat of CaV1.2 function and vascular reactivity and 2) a major risk factor for vascular complications in diabetes. A multiscale contemporary approach that includes innovative microscopy techniques, sophisticated biochemistry, electrophysiology, in silico analysis and unique animal models will be implemented to explore the following aims. Aim 1 is to elucidate the role of S1928 phosphorylation in dynamic CaV1.2 clustering and cooperative gating upon elevated glucose. Aim 2 is to examine the requirement of S1928 phosphorylation to induce CaV1.2 superclustering and cooperative gating in arterial myocytes during diabetes. Results will transform our understanding of how CaV1.2 are organized in arterial myocytes (and perhaps other cells) in health and disease and may lay the foundation for novel therapeutic strategies with single amino acid accuracy to correct channel function and vascular reactivity.

抽象的 。 给定蛋白如何组织健康和疾病的功能复合物与 用作细胞关键信息进入点的膜蛋白。一个合适且高度相关的例子 是电压门控L型CAV1.2通道，在动脉肌细胞功能和血管中起主要作用 反应性。这些通道已被证明是一致的（即合作门控），以扩大Ca2+影响。在 然而，目前对促进CAV1.2合作的机制的全面理解 门控及其在健康和疾病中的功能意义代表了主要的知识差距。总体 该提议的目的是调查要求和生理后果 血管CAV1.2通道的C末端的单个氨基酸 - S1928的磷酸化促进 CAV1.2的动态空间组织，以促进表面膜上的合作门控。完成 这个目标，我们正在测试CAV1.2 S1928磷酸化调音动态通道的中心假设 聚类和合作的门控，这有助于调节血管功能，以响应升高 细胞外葡萄糖和糖尿病期间。该假设是根据强大而严格的 初步数据揭示了S1928磷酸化作为罪魁祸首的意外和显着作用 CAV1.2亚基的重新分布和组装成动脉心肌细胞表面膜 葡萄糖和糖尿病升高。 Cav1.2由S1928磷酸化介导的超级散热器促进 CAV1.2合作门控和Ca2+影响动脉肌细胞的扩增。进一步的关键发现 强调我们观察的重要性是Cav1.2 S1928磷酸化是必要的 升高葡萄糖时的激活型信号信号通路，血管收缩和血流改变 并在糖尿病期间。此外，在新鲜分离的人动脉中已经验证了关键观察 非糖尿病患者和糖尿病患者的心肌细胞强调了转化相关性。新兴和 在本应用程序中将探讨的创新概念是S1928磷酸化的作用为1） CAV1.2功能和血管反应性的风湿病以及2）血管并发症的主要危险因素 糖尿病。一种多尺度的当代方法，包括创新的显微镜技术，精致 将实施生物化学，电生理学，电生理分析和独特的动物模型以探索 以下目标。目标1是阐明S1928磷酸化在动态CAV1.2聚类和 升高葡萄糖的合作门控。目标2是检查S1928磷酸化的要求 在糖尿病期间诱导动脉肌细胞中的CAV1.2超集群和合作门控。结果将转变 我们对CAV1.2如何在健康和其他细胞中组织的CAV1.2的理解 疾病，并可能为具有单氨基酸精度的新型治疗策略奠定基础 通道功能和血管反应性。