title: Ubiquitination of G-protein-coupled receptors.
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.
authors.affiliation: Department of Microbiology and Immunology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA.
authors.forename: Adriano
authors.initials: A
authors.lastname: Marchese
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Benovic
mesh_terms: D006801:Humans; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D043562:Receptors, G-Protein-Coupled; D025801:Ubiquitin