Metabolic Organization by the Caveolae and Cytoskeleton

描述(由申请人提供)：本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。这些代谢途径是在几十年前使用侵入性技术阐明的，因此提供了一个假设，即细胞质是统一的，本质上就像一个“酶袋”。随着具有更高空间分辨率和灵敏度的侵入性更小的技术，细胞信号通路最近被研究，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是重要的，并且细胞结构是细胞信号的关键调节成分。最近，小窝因其在细胞信号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只被粗略地研究了一下。小窝可能是糖酵解途径定位的一个部位，它允许在离子运输等耗能过程附近产生能量，并且可能对各种组织中胰岛素信号的特异性至关重要。已知细胞骨架定位于糖酵解酶，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，我们假设细胞骨架和小窝在组织和调节血管平滑肌(VSM)代谢中起重要作用。这项建议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。研究人员将测试这一假设，即微管和肌动蛋白细胞骨架作为支架将糖酵解酶固定在细胞质内(特定目标1)，以及糖酵解酶与这些结构的关联负责VSM中代谢的组织/划分。研究人员还将检验这一假设，即糖酵解酶专门定位于小窝，而糖异生酶特异定位于膜的非小窝区域(特定目标3)，以及这些特定结合负责VSM的新陈代谢组织(特定目标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，如组织代谢在支持细胞功能中的作用。

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.