Ubiquitination of G-protein-coupled receptors

Adriano Marchese; Jeffrey L Benovic

doi:10.1385/1-59259-754-8:299

Ubiquitination of G-protein-coupled receptors

Methods Mol Biol. 2004:259:299-305. doi: 10.1385/1-59259-754-8:299.

Authors

Adriano Marchese¹, Jeffrey L Benovic

Affiliation

¹ Department of Microbiology and Immunology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA.

PMID: 15250500
DOI: 10.1385/1-59259-754-8:299

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Humans
Receptors, G-Protein-Coupled / metabolism*
Ubiquitin / metabolism*

Substances

Receptors, G-Protein-Coupled
Ubiquitin