Study of GPCR–Protein Interactions Using Gel Overlay Assays and Glutathione-S-Transferase-Fusion Protein Pull-Downs
Numerous recent studies have suggested that the predicted cytosolic domains of G protein-coupled receptors represent a surface for association with proteins that may serve multiple roles in receptor localization, turnover, and signaling beyond the well-characterized interactions of these receptors with heterotrimeric G proteins. This Chapter describes two in vitro methods for ascertaining interactions between G protein-coupled receptors and various binding partners: gel overlay strategies and GST-fusion protein pull-downs.
Key wordsGel overlay G protein-coupled receptor Glutathione-S-transferase pull-down Protein–protein interaction
- 7.Hall, R. A., Ostedgaard, L. S., Fremont, R. T., et al. (1998) A C-terminal motif found in the β2-adrenergic receptor, P2Y1 receptor and cystic fibrosis transmembrane conductance regulator determines binding to the Na+/H+ exchanger regulatory factor family of PDZ proteins. Proc. Natl. Acad. Sci. USA 95, 8496–8501.PubMedCrossRefGoogle Scholar