Metabolic Organization by the Caveolae and Cytoskeleton

描述（申请人提供）：本项目将研究细胞骨架和小窝在组织和代谢调节中的作用。几十年前，使用侵入性技术阐明了代谢途径，因此提供了细胞质是均匀的并且基本上表现为“酶袋”的假设。随着具有更高空间分辨率和灵敏度的侵入性更小的技术，细胞信号传导途径最近已经被研究，并且已经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是重要的，并且细胞结构是细胞信号传导的关键调节组分。细胞膜小窝因其在细胞信号转导中的调节作用而受到广泛关注，然而，其在细胞代谢中的作用仅得到了初步研究。小窝是糖酵解途径定位的可能位点，其允许在能量消耗过程（例如离子转运）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的功能组织中的作用尚不清楚。因此，我们推测细胞骨架和小窝是重要的组织和调节血管平滑肌（VSM）代谢。这个建议的目的是确定的作用，细胞骨架（肌动蛋白和微管蛋白）和小窝结构的代谢在VSM。研究人员将测试以下假设：微管和肌动蛋白细胞骨架作为细胞质内锚定糖酵解酶的支架（具体目标1），糖酵解酶与这些结构的关联负责VSM中代谢的组织/分隔。研究人员还将检验以下假设：糖酵解酶特异性地定位于小窝，而促酵解酶特异性地定位于膜的非小窝区域（具体目标3），并且这些特异性关联负责VSM中的代谢组织（具体目标4）。这些研究可能适用于各种各样的细胞类型，并将提供一个新的理解的结构基础的组织代谢在VSM细胞和开始揭示的功能性后果的组织代谢，如有组织的代谢在支持细胞功能的作用。

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.