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Ubiquitination of G-Protein-Coupled Receptors

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Part of the Methods in Molecular Biology book series (MIMB, volume 259)

Abstract

In this chapter we describe methods for detecting the ubiquitination state of G-protein-coupled receptors (GPCRs). This involves coexpression of a GPCR with an epitope-tagged ubiquitin construct in a heterologous expression system. Modification by ubiquitin of the GPCR resulting from agonist activation is detected by immunoprecipation and subsequent immunoblotting for the epitope-tagged ubiquitin. We use here the chemokine receptor CXCR4 as the model receptor; however, this could be easily modified to detect the ubiquitination state of any GPCR.

Key Words

Agonist CXCR4 degradation G-protein-coupled receptor immunoblot immunoprecipitation lysosome sorting ubiquitin 
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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  1. 1.Department of Microbiology and Immunology, Kimmel Cancer CenterThomas Jefferson UniversityPhiladelphia

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