Study of G-protein-coupled receptor-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 (GPCRs) 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 GPCRs and various binding partners: gel overlay strategies and GST-fusion protein pull-downs.
Brady
Ashley E
AE
Vanderbilt University Medical Center, Nashville, TN, USA.
Wang
Qin
Q
Limbird
Lee E
LE
D000818:Animals; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
D043562:Receptors, G-Protein-Coupled; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase