| First Author | Shevchuk AI | Year | 2012 |
| Journal | J Cell Biol | Volume | 197 |
| Issue | 4 | Pages | 499-508 |
| PubMed ID | 22564416 | Mgi Jnum | J:185129 |
| Mgi Id | MGI:5427514 | Doi | 10.1083/jcb.201109130 |
| Citation | Shevchuk AI, et al. (2012) An alternative mechanism of clathrin-coated pit closure revealed by ion conductance microscopy. J Cell Biol 197(4):499-508 |
| abstractText | Current knowledge of the structural changes taking place during clathrin-mediated endocytosis is largely based on electron microscopy images of fixed preparations and x-ray crystallography data of purified proteins. In this paper, we describe a study of clathrin-coated pit dynamics in living cells using ion conductance microscopy to directly image the changes in pit shape, combined with simultaneous confocal microscopy to follow molecule-specific fluorescence. We find that 70% of pits closed with the formation of a protrusion that grew on one side of the pit, covered the entire pit, and then disappeared together with pit-associated clathrin-enhanced green fluorescent protein (EGFP) and actin-binding protein-EGFP (Abp1-EGFP) fluorescence. This was in contrast to conventionally closing pits that closed and cleaved from flat membrane sheets and lacked accompanying Abp1-EGFP fluorescence. Scission of both types of pits was found to be dynamin-2 dependent. This technique now enables direct spatial and temporal correlation between functional molecule-specific fluorescence and structural information to follow key biological processes at cell surfaces. |
