GPCR oligomerization and receptor trafficking

Methods Enzymol. 2013:521:69-90. doi: 10.1016/B978-0-12-391862-8.00004-1.

Abstract

The effects of oligomerization of G protein-coupled receptors (GPCRs) upon their trafficking around the cell are considerable, and this raises the potential of significant impact upon the use of existing pharmacological agents and the development of new ones. Herein, we describe a number of different techniques that can be used to study receptor dimerization/oligomerization and trafficking, beginning with a cellular system which allows the expression of two GPCRs simultaneously, one under inducible control. Subsequently, we describe means to visualize and monitor the movement of GPCRs within the cell, detect oligomerization by both resonance energy transfer and more traditional biochemical approaches, and to measure the internalization of GPCRs as part of the process of receptor regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotinylation
  • Fluorescence Resonance Energy Transfer / methods*
  • Gene Expression
  • Humans
  • Microscopy, Fluorescence / methods
  • Molecular Probe Techniques
  • Protein Multimerization
  • Protein Transport
  • Receptors, G-Protein-Coupled / analysis*
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism*

Substances

  • Receptors, G-Protein-Coupled