Monitoring receptor trafficking following retromer and WASH deregulation

Douglas G. Osborne, Christine A. Phillips-Krawczak, Daniel D. Billadeau

Research output: Chapter in Book/Report/Conference proceedingConference contribution

4 Scopus citations

Abstract

Cell surface receptors that have been internalized and enter the endocytic pathway have multiple fates including entrance into the multivesicular body pathway on their way to lysosomal degradation, recycling back to the cell surface, or retrograde trafficking out of the endolysosomal system back to the Golgi apparatus. Two ubiquitously expressed protein complexes, WASH and the endosomal coat complex retromer, function together to play a central role in directing the fate of receptors into the latter two pathways. In this chapter, we describe fluorescent- and flow cytometry-based methods for analyzing the recycling and retrograde trafficking of two receptors, α5β1 and CI-M6PR, whose intracellular fates are regulated by WASH and retromer activity. The guidelines presented in this chapter can be applied to the analysis of any cell surface or intracellular membrane protein to determine the impact of WASH or retromer deregulation on its intracellular trafficking route.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
Subtitle of host publicationSorting and Recycling Endosomes, 2015
EditorsWei Guo
PublisherAcademic Press Inc
Pages199-213
Number of pages15
ISBN (Print)9780128028292
DOIs
StatePublished - 2015

Publication series

NameMethods in Cell Biology
Volume130
ISSN (Print)0091-679X

Keywords

  • Endosome
  • F-actin
  • Receptor trafficking
  • Retromer
  • WASH

ASJC Scopus subject areas

  • Cell Biology

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