Analysis of GPCR localization and trafficking

James N Hislop, Mark von Zastrow

Research output: Chapter in Book/Report/Conference proceedingChapter

16 Citations (Scopus)

Abstract

Localization and trafficking of G protein-coupled receptors (GPCRs) is increasingly recognized to play a fundamental role in receptor-mediated signaling and its regulation. Individual receptors, including closely homologous subtypes with otherwise similar functional properties, can differ considerably in their membrane trafficking properties. In this chapter, we describe several approaches for experimentally assessing the subcellular localization and trafficking of selected GPCRs. Firstly, we describe a flexible method for receptor localization using fluorescence microscopy. We then describe two complementary approaches, using fluorescence flow cytometry and surface biotinylation, for examining receptor internalization and trafficking in the endocytic pathway.
Original languageEnglish
Title of host publicationReceptor Signal Transduction Protocols
Subtitle of host publicationThird Edition
EditorsGary B Willars, R A John Challiss
PublisherHumana Press
Pages425-440
Number of pages16
Volume746
ISBN (Electronic)978-1-61779-126-0
ISBN (Print)978-1-61779-125-3
DOIs
Publication statusPublished - 2011

Publication series

NameMethods in Molecular Biology: Methods and Protocols
Volume746
ISSN (Print)1064-3745

Keywords

  • biotinylation
  • flow cytometry
  • HEK293 cells
  • humans
  • intracellular space
  • protein transport
  • receptors, G-protein-coupled
  • staining and labeling
  • tissue fixation

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  • Cite this

    Hislop, J. N., & von Zastrow, M. (2011). Analysis of GPCR localization and trafficking. In G. B. Willars, & R. A. J. Challiss (Eds.), Receptor Signal Transduction Protocols: Third Edition (Vol. 746, pp. 425-440). (Methods in Molecular Biology: Methods and Protocols; Vol. 746). Humana Press. https://doi.org/10.1007/978-1-61779-126-0_25