Metabolic Organization by the Caveolae and Cytoskeleton

小凹和细胞骨架的代谢组织

• 批准号: 6614744
• 负责人: Christopher D Hardin
• 金额: $ 23.78万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6614744
• 关键词: SDS polyacrylamide gel electrophoresis; actins; caveolins; confocal scanning microscopy; cytoskeleton; electron microscopy; enzymes; gluconeogenesis; glycolysis; immunoprecipitation; membrane proteins; metabolism; microtubules; nuclear magnetic resonance spectroscopy; vascular smooth muscle; western blottings

DESCRIPTION (provided by applicant): This project will examine the role of the cytoskeleton and the caveolae in the organization and regulation of metabolism. The metabolic pathways were elucidated decades ago using invasive techniques and thus provided an assumption that the cytoplasm was uniform and essentially behaved as a "bag of enzymes". With less invasive techniques with higher spatial resolution and sensitivity, cell signaling pathways have recently been studied and a very different paradigm has arisen that cytoarchitecture is important for localization of signaling proteins and small molecules and that the cytoarchitecture is a key regulatory component for cell signaling. The caveolae have recently received considerable attention for their role in the regulation of cell signaling, however, their role in cell metabolism has been only superficially investigated. Caveolae are a likely site of glycolytic pathway localization which allows for the production of energy near energy-consuming processes such as ion transport and are likely critical to the specificity of insulin signaling in a variety of tissues. The cytoskeleton has been known to localize glycolytic enzymes but the role of such localization in the functional organization of glycolytic metabolism is not known. Therefore, we hypothesize that the cytoskeleton and the caveolae are important in the organization and regulation of vascular smooth muscle (VSM) metabolism. The aim of this proposal is to determine the role of the cytoskeleton (actin and tubulin) and caveolae in the structuring of metabolism in VSM. The investigators will test the hypothesis that microtubules and the actin cytoskeleton function as a scaffolding for anchoring glycolytic enzymes within the cytoplasm (specific aim 1) and that the association of glycolytic enzymes with these structures is responsible for the organization/compartmentation of metabolism in VSM. The investigators will also test the hypothesis that glycolytic enzymes are specifically localized to caveolae while gluconeogenic enzymes are specifically localized to non-caveolar regions of the membrane (specific aim 3) and that these specific associations are responsible for the organization of metabolism in VSM (specific aim 4). These investigations are likely applicable to a wide variety of cell types and will provide a new understanding of the structural basis for the organization of metabolism in VSM cells and begin to unveil the functional consequences of the organization of metabolism such as the role of organized metabolism in the support of cell function.

描述（由申请人提供）：该项目将研究细胞骨架和小凹在新陈代谢的组织和调节中的作用。几十年前利用侵入性技术阐明了代谢途径，从而提供了细胞质是均匀的并且本质上表现为“酶袋”的假设。利用具有更高空间分辨率和灵敏度的侵入性较小的技术，最近对细胞信号传导途径进行了研究，并出现了一种非常不同的范式，即细胞结构对于信号蛋白和小分子的定位非常重要，并且细胞结构是细胞信号传导的关键调节成分。小凹最近因其在细胞信号传导调节中的作用而受到相当大的关注，然而，它们在细胞代谢中的作用仅得到了肤浅的研究。小凹可能是糖酵解途径定位的位点，它允许在离子运输等能量消耗过程附近产生能量，并且可能对多种组织中胰岛素信号传导的特异性至关重要。已知细胞骨架定位糖酵解酶，但这种定位在糖酵解代谢功能组织中的作用尚不清楚。因此，我们假设细胞骨架和小凹在血管平滑肌（VSM）代谢的组织和调节中很重要。该提案的目的是确定细胞骨架（肌动蛋白和微管蛋白）和小凹在 VSM 代谢结构中的作用。研究人员将检验以下假设：微管和肌动蛋白细胞骨架充当将糖酵解酶锚定在细胞质内的支架（具体目标 1），并且糖酵解酶与这些结构的关联负责 VSM 中代谢的组织/区室。研究人员还将检验以下假设：糖酵解酶专门定位于膜凹，而糖异生酶专门定位于膜的非凹凸区域（具体目标 3），并且这些特定关联负责 VSM 中的代谢组织（具体目标 4）。这些研究可能适用于多种细胞类型，并将为 VSM 细胞代谢组织的结构基础提供新的理解，并开始揭示代谢组织的功能后果，例如有组织的代谢在支持细胞功能中的作用。