Study of GPCR-protein interactions using gel overlay assays and glutathione-S-transferase-fusion protein pull-downs.

Ashley E Brady, Yunjia Chen, Lee E Limbird, Qin Wang

Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN, USA.

Methods in molecular biology (Clifton, N.J.) 2011

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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.

Citation

Ashley E Brady, Yunjia Chen, Lee E Limbird, Qin Wang. Study of GPCR-protein interactions using gel overlay assays and glutathione-S-transferase-fusion protein pull-downs. Methods in molecular biology (Clifton, N.J.). 2011;746:347-55