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: 10306953
• 负责人: Manuel F Navedo
• 金额: $ 57.91万
• 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/10306953
• 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 。 特定蛋白质如何在健康和疾病中组织成功能复合物与以下因素特别相关： 膜蛋白作为细胞的关键信息入口点。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功能中起主要作用 反应性。这些通道已被证明可以一致门控（即合作门控）以放大 Ca2+ 流入。在 然而，目前对促进 CaV1.2 合作诱导机制的全面了解 门控及其对健康和疾病的功能影响代表了主要的知识差距。整体 该提案的目的是调查需求和生理后果，从而 血管 CaV1.2 通道 C 端单个氨基酸 – S1928 的磷酸化促进 CaV1.2 的动态空间组织促进表面膜的协同门控。为了完成 为了这个目标，我们正在测试 CaV1.2 S1928 磷酸化调节动态通道的中心假设 聚类和协作门控，这有助于调节血管功能以响应升高的 细胞外葡萄糖和糖尿病期间。这个假设是建立在强有力的、严格的基础上的。 初步数据揭示了 S1928 磷酸化作为罪魁祸首的意想不到的显着作用 CaV1.2 亚基重新分布并组装成动脉肌细胞表面膜的超簇 血糖升高和糖尿病。 S1928磷酸化介导的CaV1.2超簇促进 CaV1.2 协同门控和 Ca2+ 流入动脉肌细胞的放大。主要发现进一步 强调我们观察的重要性是 CaV1.2 S1928 磷酸化对于 血糖升高时激活降压信号通路、血管收缩和血流改变 以及糖尿病期间。此外，关键观察结果已在新鲜分离的人类动脉中得到验证。 来自非糖尿病和糖尿病患者的肌细胞，强调了转化相关性。新兴和 本应用中将探讨的创新概念是 S1928 磷酸化的作用：1) CaV1.2 功能和血管反应性的变阻器以及 2) 血管并发症的主要危险因素 糖尿病。多尺度当代方法，包括创新的显微镜技术、复杂的 将采用生物化学、电生理学、计算机分析和独特的动物模型来探索 以下目标。目标 1 是阐明 S1928 磷酸化在动态 CaV1.2 聚类和 葡萄糖升高时的合作门控。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 超聚类和协作门控。结果将会改变 我们对 CaV1.2 在健康和健康状况下的动脉肌细胞（或许还有其他细胞）中如何组织的理解 疾病，并可能为具有单氨基酸准确性的新治疗策略奠定基础，以纠正 通道功能和血管反应性。