Analyzing the role of AP-1B in polarized sorting from recycling endosomes in epithelial cells

Heike Fölsch*

*Corresponding author for this work

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

2 Scopus citations

Abstract

Epithelial cells polarize their plasma membrane into apical and basolateral domains where the apical membrane faces the luminal side of an organ and the basolateral membrane is in contact with neighboring cells and the basement membrane. To maintain this polarity, newly synthesized and internalized cargos must be sorted to their correct target domain. Over the last ten years, recycling endosomes have emerged as an important sorting station at which proteins destined for the apical membrane are segregated from those destined for the basolateral membrane. Essential for basolateral sorting from recycling endosomes is the tissue-specific adaptor complex AP-1B. This chapter describes experimental protocols to analyze the AP-1B function in epithelial cells including the analysis of protein sorting in LLC-PK1 cells lines, immunoprecipitation of cargo proteins after chemical crosslinking to AP-1B, and radioactive pulse-chase experiments in MDCK cells depleted of the AP-1B subunit μ1B.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
Subtitle of host publicationSorting and Recycling Endosomes, 2015
EditorsWei Guo
PublisherAcademic Press Inc
Pages289-305
Number of pages17
ISBN (Print)9780128028292
DOIs
StatePublished - Jan 1 2015

Publication series

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

Keywords

  • AP-1B
  • Basolateral membrane
  • Epithelial cell
  • LLC-PK1
  • MDCK
  • Polarized sorting
  • Recycling endosome

ASJC Scopus subject areas

  • Cell Biology

Fingerprint Dive into the research topics of 'Analyzing the role of AP-1B in polarized sorting from recycling endosomes in epithelial cells'. Together they form a unique fingerprint.

Cite this