Green Fluorescent Protein-Tagged β-Arrestin Translocation as a Measure of G Protein-Coupled Receptor Activation

  • Stephen S. G. Ferguson
  • Marc G. Caron
Part of the Methods in Molecular Biology™ book series (MIMB, volume 237)


The G protein-coupled receptor (GPCR) superfamily is the largest family of integral membrane proteins. GPCRs respond to a wide variety of sensory and chemical stimuli and contribute directly to the regulation of all major organ systems. As such, GPCRs represent primary drug targets for therapeutic intervention. Although GPCRs respond to a diverse range of ligands and signal through multiple heterotrimeric G proteins, the inactivation of GPCR signaling is mediated by a limited set of proteins. In particular, the desensitization of the majority of GPCRs is mediated by the binding of two arrestin isoforms, β-arrestin1 and β-arrestin2, that exhibit overlapping substrate specificity. In response to GPCR activation and phosphorylation by GPCR kinases, β-arrestins redistribute from the cytosol to the plasma membrane to bind GPCRs and subsequently target the receptors for internalization via clathrin-coated vesicles. This property of β-arrestins has allowed the development of a green fluorescent protein (GFP)-based assay for detecting GPCR activation by confocal microscopy. This β-arrestin-GFP translocation methodology is described in detail in this chapter.

Key Words

β-arrestin green fluorescent protein G protein-coupled receptor G protein-coupled receptor kinase confocal microscopy translocation clathrin dynamin endocytosis agonist antagonist plasma membrane 


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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  • Stephen S. G. Ferguson
    • 1
  • Marc G. Caron
    • 2
  1. 1.The John P. Robarts Institute and Departments of Physiology, Pharmacology and Toxicology, and MedicineUniversity of Western OntarioLondonCanada
  2. 2.Howard Hughes Medical InstituteDuke University Medical CenterDurham

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