Скачать или смотреть Morphological changes of plasma membrane and protein assembly during clathrin-mediated endocytosis

Title:
Morphological changes of plasma membrane and protein assembly during clathrin-mediated endocytosis
Author:
Aiko Yoshida, Nobuaki Sakai, Yoshitsugu Uekusa, Yuka Imaoka, Yoshitsuna Itagaki, Yuki Suzuki, Shige H. Yoshimura
Published in:
PLOS Biol 16(5): e2004786.
https://doi.org/10.1371/journal.pbio....

Research Report
Graduate School of Biostudies, Kyoto University, Kyoto, Japan
Laboratory of Plasma Membrane and Nuclear Signaling,
Ph.D, Aiko Yoshida

研究成果報告
京都大学大学院　生命科学研究科　統合生命科学専攻
分子情報解析学分野　吉村研究室
元博士後期課程学生（現北海道大学博士研究員）吉田藍子

Abstract:
Clathrin-mediated endocytosis (CME) proceeds through a series of morphological changes of the plasma membrane induced by a number of protein components. Although the spatiotemporal assembly of these proteins has been elucidated by fluorescence-based techniques, the protein-induced morphological changes of the plasma membrane have not been fully clarified in living cells. Here, we visualize membrane morphology together with protein localizations during CME by utilizing high-speed atomic force microscopy (HS-AFM) combined with a confocal laser scanning unit. The plasma membrane starts to invaginate approximately 30 s after clathrin starts to assemble, and the aperture diameter increases as clathrin accumulates. Actin rapidly accumulates around the pit and induces a small membrane swelling, which, within 30 s, rapidly covers the pit irreversibly. Inhibition of actin turnover abolishes the swelling and induces a reversible open–close motion of the pit, indicating that actin dynamics are necessary for efficient and irreversible pit closure at the end of CME.