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Rab GTPases pp 283-293 | Cite as

Role of the Rab5 Guanine Nucleotide Exchange Factor, Rme-6, in the Regulation of Clathrin-Coated Vesicle Uncoating

  • Elizabeth SmytheEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1298)

Abstract

Clathrin-coated pits are major ports of entry into eukaryotic cells. Following scission of clathrin-coated pits to form coated vesicles, the peripheral membrane proteins that form the coat need to be removed. Here we describe an assay that provides a measure of the extent of clathrin coat uncoating in intact cells. This assay has been used to investigate rab5-dependent modulation of uncoating. Specifically, it has been used to identify functional differences between rab5 guanine nucleotide exchange factors.

Key words

Endocytosis rab5 Guanine nucleotide exchange factors Rme-6 AP2 Clathrin Coated vesicles Uncoating 

Notes

Acknowledgments

Research in the author’s lab is supported by the MRC, the Wellcome Trust and Cancer Research UK. Images were collected at the Light Microscopy Facility at the University of Sheffield.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Biomedical Science, Centre for Membrane Interactions and DynamicsUniversity of SheffieldSheffieldUK

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