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  • Title: Bioluminescence Resonance Energy Transfer (BRET) to Detect the Interactions Between Kappa Opioid Receptor and Nonvisual Arrestins.
    Author: Bedini A.
    Journal: Methods Mol Biol; 2021; 2201():45-58. PubMed ID: 32975788.
    Abstract:
    Bioluminescence resonance energy transfer (BRET ) is a very sensitive technique employed to study protein-protein interactions, including G-protein-coupled receptor (GPCR ) hetero- and homo-dimerization. Recently, BRET has also been used to investigate the interaction between GPCRs (e.g.: α2 adrenergic receptor, muscarinic M2 receptor, dopaminergic D2 receptor) and nonvisual arrestins. Within the last decade an increasing interest arose toward opioid agonists with limited activation of arrestin-dependent signaling pathways, as they are believed to be effective analgesics with reduced adverse effects. Here a BRET protocol is described to investigate interactions between the kappa opioid receptor (KOR ) and nonvisual arrestins (arrestin-2 and arrestin-3) in HEK-293 cells, both under basal conditions and after exposure to KOR ligands.
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