Structural basis for caveolae assembly and function

小窝组装和功能的结构基础

• 批准号: 9925038
• 负责人: Anne K Kenworthy
• 金额: $ 52.77万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9925038
• 关键词: Acute Lung Injury; Address; Architecture; Asthma; Binding; Biochemical; Biogenesis; Biological Assay; Biology; Blood Vessels; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Caveolae; Cell membrane; Cell physiology; Cells; Chronic; Complex; Computer Simulation; Cryoelectron Microscopy; Data; Defect; Dimensions; Disease; Electron Microscopy; Ensure; Fluorescence Polarization; Fluorescence Spectroscopy; Goals; Hand; Homeostasis; Inflammatory; Knowledge; Link; Lipids; Literature; Lung; Lung diseases; Malignant neoplasm of lung; Maps; Membrane; Membrane Proteins; Modeling; Mutation; Protein Region; Proteins; Pulmonary Fibrosis; Research Personnel; Resolution; Shapes; Signal Transduction; Structure; Surface; System; Testing; Variant; caveolin 1; cell type; flasks; insight; monomer; personalized medicine; pulmonary arterial hypertension; pulmonary function; single molecule; stoichiometry; three dimensional structure; trafficking

Flask-shaped invaginations of the plasma membrane known as caveolae are important regulators of the cardiovascular and pulmonary systems. The membrane protein caveolin-1 (Cav1) is a major structural component of caveolae and is required for their formation in non-muscle cells. Cav1 is highly expressed in the lung, and has been linked to lung cancer, asthma, pulmonary fibrosis, pulmonary arterial hypertension, chronic inflammatory respiratory diseases, and acute lung injury. Cav1 is also known to regulate vascular homeostasis and was recently identified as one of 15 key drivers of cardiovascular disease. However, the exact mechanisms by which caveolae form and function normally, and how defects in caveolae give rise to disease remain incompletely understood. One of the greatest impediments to our understanding of how caveolae assemble and function is our limited knowledge about how Cav1 is packed within caveolae. The goal of this proposal is to address this major gap in knowledge by defining the atomic-level structure of Cav1 oligomers that are known to serve as the fundamental building blocks of caveolae. To do so, we will utilize a combination of biochemical and spectroscopic approaches, single molecule electron microscopy, cell biological assays, and computational modeling to study purified Cav1 oligomeric complexes. One specific aim is to define the overall architecture of Cav1 oligomers and identify key determinants of their structure and stoichiometry. Another aim is to map the three-dimensional organization of Cav1 within these oligomers and define structural changes in the protein associated with the monomer to oligomer transition. The results of these studies will enable us to answer a number of long-standing questions in the field, including how Cav1 monomers oligomerize to form complexes, what structural features of Cav1 are required for the protein to bend membranes, how Cav1 complexes interact amongst themselves to build caveolae, and what regions of Cav1 are available to bind other proteins and lipids. These insights will be key to furthering our understanding of how Cav1 and caveolae regulate cellular functions that are critical to ensure proper function of the cardiovascular and pulmonary systems. Finally, our studies will also have important implications for personalized medicine by providing a structural framework for understanding how both common variants and disease-associated mutations of Cav1 impact the structure and function of caveolae. !

被称为小窝的质膜的烧瓶状内陷是细胞膜的重要调节因子。 心血管和肺系统。膜蛋白Caveolin-1（Cav 1）是一种主要的结构蛋白， 细胞膜是细胞膜小窝的组成部分，是它们在非肌肉细胞中形成所必需的。Cav 1在人的大脑中高度表达， 肺，并已与肺癌，哮喘，肺纤维化，肺动脉高压，慢性 炎性呼吸道疾病和急性肺损伤。已知Cav 1还调节血管稳态 最近被确定为心血管疾病的15个关键驱动因素之一。但具体 小窝形成和功能正常的机制，以及小窝缺陷如何引起疾病 仍然不完全理解。 我们理解小窝如何组装和发挥功能的最大障碍之一是我们有限的 了解Cav 1是如何在小窝中包装的。该提案的目标是解决这一重大差距 通过定义已知作为分子的Cav 1低聚物的原子级结构， 是构成洞穴的基础为此，我们将利用生物化学和 光谱方法，单分子电子显微镜，细胞生物学测定，和计算 建模以研究纯化的Cav 1寡聚复合物。一个具体的目标是定义 Cav 1寡聚体，并确定其结构和化学计量的关键决定因素。另一个目标是绘制 Cav 1在这些寡聚体中的三维组织，并定义蛋白质中的结构变化 与单体到低聚物的转变有关。这些研究的结果将使我们能够回答一个 该领域的一些长期存在的问题，包括Cav 1单体如何寡聚形成复合物， Cav 1的什么结构特征是蛋白质弯曲膜所必需的，Cav 1复合物如何相互作用 以及Cav 1的哪些区域可用于结合其他蛋白质， 脂质。这些见解将是进一步了解Cav 1和caveolae如何调节细胞的关键。 这些功能对于确保心血管和肺系统的正常功能至关重要。最后我们 研究还将通过提供以下结构框架，对个性化医疗产生重要影响： 了解Cav 1的常见变异和疾病相关突变如何影响结构， 小窝的功能。 !