De novo endocytic clathrin coats develop curvature at early stages of their formation
De novo endocytic clathrin coats develop curvature at early stages of their formation
复制标题
DOI：
10.1016/j.devcel.2021.10.019
复制
发表时间：
2021-11-22
影响因子：
11.8
通讯作者：
Kural, Comert
中科院分区：
文献类型：
--
作者：
Willy, Nathan M.;Ferguson, Joshua P.;Kural, Comert
将一个平坦的膜片雕刻成内吞囊泡需要在细胞表面产生曲率，这是内吞机制的主要功能。利用超分辨活细胞荧光成像，我们证明了单个网格蛋白包被凹坑产生的曲率可以在培养的细胞和发育中的生物组织中实时检测到。我们的分析表明，在不同的质膜张力水平下，网格蛋白外壳的足迹在形成坑的过程中单调增加。这些发现仅与预测内吞网格蛋白坑形成早期阶段曲率产生的模型相一致。我们还发现，与网格蛋白斑块相关的CALM接头形成簇状，而AP2的分布更为均匀。考虑到CALM的曲率感知和驱动作用，我们提出CALM簇可能会局部增加网格蛋白晶格上的应变，最终导致斑块边缘的破裂和随后的凹坑完成。
Sculpting a flat patch of membrane into an endocytic vesicle requires curvature generation on the cell surface, which is the primary function of the endocytosis machinery. Using super-resolved live cell fluorescence imaging, we demonstrate that curvature generation by individual clathrin-coated pits can be detected in real time within cultured cells and tissues of developing organisms. Our analyses demonstrate that the footprint of clathrin coats increases monotonically during the formation of pits at different levels of plasma membrane tension. These findings are only compatible with models that predict curvature generation at the early stages of endocytic clathrin pit formation. We also found that CALM adaptors associated with clathrin plaques form clusters, whereas AP2 distribution is more homogenous. Considering the curvature sensing and driving roles of CALM, we propose that CALM clusters may increase the strain on clathrin lattices locally, eventually giving rise to rupture and subsequent pit completion at the edges of plaques.