Structural basis for caveolae assembly and function

烧瓶状的质膜内陷称为小窝，是调节细胞周期的重要因素。 心血管和肺脏系统。膜蛋白小窝蛋白-1(Cav1)是一种主要的结构蛋白 小窝的组成部分，是在非肌肉细胞中形成小窝所必需的。Cav1在细胞中高度表达 肺，并与肺癌、哮喘、肺纤维化、肺动脉高压、慢性 炎症性呼吸道疾病和急性肺损伤。Cav1也是已知的调节血管内稳态的基因。 最近被确定为心血管疾病的15个关键驱动因素之一。然而，准确的 小窝正常形成和功能的机制，以及小窝缺陷如何导致疾病 仍然没有完全被理解。 我们理解小窝如何组装和功能的最大障碍之一是我们有限的 关于Cav1是如何在小窝中填充的知识。这项提案的目标是解决这一重大差距 通过定义Cav1低聚物的原子级结构，这些低聚物被认为是 洞穴的基本构件。要做到这一点，我们将利用生化和 光谱方法、单分子电子显微镜、细胞生物学分析和计算 用于研究纯化的Cav1寡聚复合体的建模。一个特定的目标是定义的总体架构 Cav1低聚物，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 Cav1在这些寡聚体中的三维组织和定义蛋白质的结构变化 伴随着单体向齐聚物的转变。这些研究的结果将使我们能够回答 该领域的一些长期存在的问题，包括Cav1单体如何齐聚形成络合物， Cav1的哪些结构特征是蛋白质弯曲细胞膜所必需的，Cav1复合体如何相互作用 它们之间建立小窝，以及Cav1的哪些区域可用于结合其他蛋白质和 脂类。这些见解将是进一步理解Cav1和Caveolae如何调控细胞的关键 对确保心血管和肺脏系统的正常功能至关重要的功能。最后，我们的 研究还将为个性化医疗提供一个结构框架，从而对个性化医疗产生重要影响 了解Cav1的常见变异和疾病相关突变如何影响结构和 小窝的功能。 好了！

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. !