TY - JOUR T1 - Interaction Interface between 5-HT3A serotonin receptor and RIC-3 chaperone JF - bioRxiv DO - 10.1101/2022.02.17.480943 SP - 2022.02.17.480943 AU - Hoa Quynh Do AU - Michaela Jansen Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/02/19/2022.02.17.480943.abstract N2 - Serotonin or 5-hydroxytryptamine receptors type 3 (5-HT3) belong to the family of pentameric ligand-gated ion channels (pLGICs), which also includes other neurotransmitter-gated ion channels such as nicotinic acetylcholine receptors, γ-amino butyric acid A receptors, and glycine receptors. pLGICs have been long-standing clinical targets for treating psychiatric disorders such as anxiety, schizophrenia, addiction, and neurological diseases like Alzheimer’s and Parkinson’s disease. Due to the highly-homologous structures of pLGICs’ extracellular and transmembrane domains, clinical trials for drug candidates targeting these domains have been largely hampered by undesired effects mediated by off-subunit modulation. With the present study, we explore the interaction interface of the 5-HT3A intracellular domain (ICD) with the resistance to inhibitors of choline esterase (RIC-3) chaperone. Protein-protein interactions are attractive and successful targets for contemporary drug development. Previously, we have shown that RIC-3 physically interacts with 5-HT3A subunits, and we identified the L1-MX fragment of the ICD fused to maltose-binding protein as sufficient for interaction. For the present study, synthetic L1-MX-based fragments and Ala-scanning were used to probe binding to RIC-3 using a pull-down assay. In complementary studies with full-length 5-HT3A subunits, we investigated the impact of these substitutions on modulation of functional surface expression by RIC-3 using two-electrode voltage-clamp recording after expression in Xenopus laevis oocytes. Our data demonstrate that the 5-HT3A-MX segment contains the interaction interface, and that residues W347, R349, and L353 are critical for the interaction with RIC-3.Competing Interest StatementThe authors have declared no competing interest.5-HT3The serotonin receptor type 3nAChRnicotinic acetylcholine receptorpLGICspentameric ligand-gated ion channelsPPIprotein-protein interaction ER -