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Updated: Jan 26, 2026

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies
Published on: July 24, 2017
Combining SRET2 and BiFC to Study GPCR Heteromerization and Protein-Protein Interactions
Amina M Bagher1,2, Melanie E M Kelly1,3, Eileen M Denovan-Wright4
1Department of Pharmacology, Dalhousie University, Halifax, NS, Canada.
Abstract:
G protein-coupled receptors (GPCRs) are the target for many drugs. Evidence continues to accumulate demonstrating that multiple receptors form homo- and heteromeric complexes, which in turn dynamically couple with G proteins, and other interacting proteins. Here, we describe a method to simultaneously determine the identity of up to four distinct constituents of GPCR complexes using a combination of sequential bioluminescence resonance energy transfer 2-fluorescence resonance energy transfer (SRET2) with bimolecular fluorescence complementation (BiFC). The method is amenable to moderate throughput screening of changes in response to ligands and time-course analysis of protein-protein oligomerization.
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