Preferential Gs protein coupling of the galanin Gal1 receptor in the µ-opioid-Gal1 receptor heterotetramer. (August 2022)

• Record Type: Journal Article
• Authors: De Oliveira, Paulo A.; Moreno, Estefanía; Casajuana-Martin, Nil; Casadó-Anguera, Verònica; Cai, Ning-Sheng; Camacho-Hernandez, Gisela Andrea; Zhu, Hu; Bonifazi, Alessandro; Hall, Matthew D.; Weinshenker, David; Newman, Amy Hauck; Logothetis, Diomedes E.; Casadó, Vicent; Plant, Leigh D.; Pardo, Leonardo; Ferré, Sergi
• Is Part Of: Pharmacological research. Volume 182(2022)
• Publication Date: 2022-08
• Subjects: DAMGO (PubChem CID: 5462471) -- Endomorphin-1 (PubChem CID: 5311080) -- Galanin (PubChem CID: 16133823) -- M40 (PubChem CID: 16133821) -- M617 (PubChem CID: 16158157) -- Naloxone (PubChem CID: 5284596) -- Methadone (PubChem CID: 4095) -- Fentanyl (PubChem CID: 3345); A2AR adenosine A2A receptor -- AC adenylyl cyclase -- ANOVA analysis of variance -- BiFC bimolecular fluorescence complementation -- BRET bioluminescence resonance energy transfer -- cAMP cyclic adenosine monophosphate -- CODA-RET complemented donor-acceptor resonance energy transfer -- cryo-EM cryo-electron microscopy -- D2R dopamine D2 receptor -- DMEM Dulbecco's modified Eagle's medium -- EYFP enhanced yellow variant of GFP -- FBS fetal bovine serum -- Gal1R galanin Gal1 receptor -- FRET fluorescence resonance energy transfer -- GPCR G protein-coupled receptor -- HEK cells human embryonic kidney cell line -- mC monomeric Cherry -- mT monomeric teal fluorescent protein 1 -- mVenus monomeric Venus -- MOR µ-opioid receptors -- MU cells cell line expressing µ-opioid receptors -- MU-GAL cells cell line expressing µ-opioid receptors and galanin Gal1 receptors -- PEI polyethylenimine -- RLuc Renilla Luciferase -- TIRF total internal reflection fluorescence -- TM transmembrane domain -- VTA ventral tegmental area -- WT wild-type -- YFP yellow fluorescence protein; Opioid receptors -- Galanin receptors -- G protein coupled receptor oligomerization -- Total internal reflection fluorescence microscopy; Pharmacology -- Periodicals; Pharmacology -- Periodicals; Research -- Periodicals; Médicaments -- Recherche -- Périodiques; Pharmacologie -- Périodiques; 615.105
• DOI: 10.1016/j.phrs.2022.106322
• Languages: English
• ISSNs: 1043-6618

Abstract

Abstract: Recent studies have proposed that heteromers of µ-opioid receptors (MORs) and galanin Gal1 receptors (Gal1 Rs) localized in the mesencephalon mediate the dopaminergic effects of opioids. The present study reports converging evidence, using a peptide-interfering approach combined with biophysical and biochemical techniques, including total internal reflection fluorescence microscopy, for a predominant homodimeric structure of MOR and Gal1 R when expressed individually, and for their preference to form functional heterotetramers when co-expressed. Results show that a heteromerization-dependent change in the Gal1 R homodimeric interface leads to a switch in G-protein coupling from inhibitory Gi to stimulatory Gs proteins. The MOR-Gal1 R heterotetramer, which is thus bound to Gs via the Gal1 R homodimer and Gi via the MOR homodimer, provides the framework for a canonical Gs-Gi antagonist interaction at the adenylyl cyclase level. These novel results shed light on the intense debate about the oligomeric quaternary structure of G protein-coupled receptors, their predilection for heteromer formation, and the resulting functional significance. Graphical Abstract: ga1