An alternative mechanism of clathrin-coated pit closure revealed by ion conductance microscopy

• DOI: 10.1083/jcb.201109130
• Publication Date: 2012-05-08T06:51:53Z
• Authors: Andrew I. Shevchuk; Pavel Novak; Marcus Taylor; Ivan A. Diakonov; Azza Ziyadeh-Isleem; Marc Bitoun; Pascale Guicheney; Max J. Lab; Julia Gorelik; Christien J. Merr...; David Klenerman; Yuri E. Korchev
• Keywords: Cell membrane

Current knowledge of the structural changes taking place during clathrin-mediated endocytosis is largely based on electron microscopy images fixed preparations and x-ray crystallography data purified proteins. In this paper, we describe a study clathrin-coated pit dynamics in living cells using ion conductance to directly image shape, combined with simultaneous confocal follow molecule-specific fluorescence. We find that 70% pits closed formation protrusion grew one side pit, covered entire then disappeared together pit-associated clathrin–enhanced green fluorescent protein (EGFP) actin-binding protein–EGFP (Abp1-EGFP) This was contrast conventionally closing cleaved from flat membrane sheets lacked accompanying Abp1-EGFP Scission both types found be dynamin-2 dependent. technique now enables direct spatial temporal correlation between functional fluorescence information key biological processes at cell surfaces.