Medical College of Wisconsin
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Ubiquitination of GPCRs. Methods Mol Biol 2011;746:251-9

Date

05/25/2011

Pubmed ID

21607861

Pubmed Central ID

PMC4699430

DOI

10.1007/978-1-61779-126-0_13

Scopus ID

2-s2.0-80052791836   5 Citations

Abstract

In this chapter, we describe a method for detecting the ubiquitination status of G protein-coupled receptors (GPCRs). This involves co-expression of a GPCR with an epitope-tagged ubiquitin construct in a -heterologous mammalian expression system. Stimulus-dependent modification of the GPCR by -ubiquitin is detected by immunoprecipitation and subsequent immunoblotting to detect incorporation of the epitope-tagged ubiquitin. We describe here a well-established protocol to detect ubiquitination of the chemokine receptor CXCR4, which can be easily applied to detect the ubiquitination status of other GPCRs.

Author List

Caballero A, Marchese A

Author

Adriano Marchese PhD Professor in the Biochemistry department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

HEK293 Cells
Humans
Receptors, CXCR4
Receptors, G-Protein-Coupled
Transfection
Ubiquitin
Ubiquitination