Ubiquitination of GPCRs. Methods Mol Biol 2011;746:251-9
Date
05/25/2011Pubmed ID
21607861Pubmed Central ID
PMC4699430DOI
10.1007/978-1-61779-126-0_13Scopus ID
2-s2.0-80052791836 (requires institutional sign-in at Scopus site) 5 CitationsAbstract
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 AAuthor
Adriano Marchese PhD Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
HEK293 CellsHumans
Receptors, CXCR4
Receptors, G-Protein-Coupled
Transfection
Ubiquitin
Ubiquitination