Ubiquitination of GPCRs

Methods Mol Biol. 2011:746:251-9. doi: 10.1007/978-1-61779-126-0_13.


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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • HEK293 Cells
  • Humans
  • Receptors, CXCR4 / metabolism
  • Receptors, G-Protein-Coupled / metabolism*
  • Transfection
  • Ubiquitin / metabolism
  • Ubiquitination*


  • Receptors, CXCR4
  • Receptors, G-Protein-Coupled
  • Ubiquitin