Caveolae are small coated plasma membrane invaginations with diverse functions. Caveolae undergo curvature changes. Yet, it is unclear which proteins regulate this process. To address this gap, we develop a correlative stimulated emission depletion (STED) fluorescence and platinum replica electron microscopy imaging (CLEM) method to image proteins at single caveolae. Caveolins and cavins are found at all caveolae, independent of curvature. EHD2 is detected at both low and highly curved caveolae. Pacsin2 associates with low curved caveolae and EHBP1 with mostly highly curved caveolae. Dynamin is absent from caveolae. Cells lacking dynamin show no substantial changes to caveolae, suggesting that dynamin is not directly involved in caveolae curvature. We propose a model where caveolins, cavins, and EHD2 assemble as a cohesive structural unit regulated by intermittent associations with pacsin2 and EHBP1. These coats can flatten and curve to enable lipid traffic, signaling, and changes to the surface area of the cell.

小凹是具有多种功能的小涂层质膜内陷。小凹经历曲率变化。然而，尚不清楚哪些蛋白质调节这一过程。为了解决这一差距，我们开发了一种相关受激发射损耗（STED）荧光和铂复制电子显微镜成像（CLEM）方法来对单个小窝的蛋白质进行成像。所有小凹处都存在小凹和小凹，与曲率无关。 EHD2 在低弯曲和高弯曲的凹坑中均可检测到。 Pacsin2 与低弯曲的小窝相关，而 EHBP1 与大部分高度弯曲的小窝相关。小窝中不存在动力蛋白。缺乏动力的细胞显示小凹没有实质性变化，这表明动力不直接参与小凹曲率。我们提出了一个模型，其中 Caveolins、cavins 和 EHD2 组装为一个内聚结构单元，受 pacsin2 和 EHBP1 的间歇性关联调节。这些涂层可以变平和弯曲，以实现脂质运输、信号传导和细胞表面积的变化。