Morató Arús, XavierFernández Dueñas, VíctorPérez-Villamor, PilarValle León, MartaVela, José MiguelMerlos Roca, ManuelBurgueño Hurtado, JavierCiruela Alférez, Francisco2023-07-142023-07-142023-12-011948-7193https://hdl.handle.net/2445/200655The σ1 receptor (S1R) is a ligand-regulated non-opioid intracellular receptor involved in several pathological conditions. The development of S1R-based drugs as therapeutic agents is a challenge due to the lack of simple functional assays to identify and classify S1R ligands. We have developed a novel nanoluciferase binary technology (NanoBiT) assay based on the ability of S1R to heteromerize with the binding immunoglobulin protein (BiP) in living cells. The S1R-BiP heterodimerization biosensor allows for rapid and accurate identification of S1R ligands by monitoring the dynamics of association-dissociation of S1R and BiP. Acute treatment of cells with the S1R agonist PRE-084 produced rapid and transient dissociation of the S1R-BiP heterodimer, which was blocked by haloperidol. The effect of PRE-084 was enhanced by calcium depletion, leading to a higher reduction in heterodimerization even in the presence of haloperidol. Prolonged incubation of cells with S1R antagonists (haloperidol, NE-100, BD-1047, and PD-144418) increased the formation of S1R-BiP heteromers, while agonists (PRE-084, 4-IBP, and pentazocine) did not alter heterodimerization under the same experimental conditions. The newly developed S1R-BiP biosensor is a simple and effective tool for exploring S1R pharmacology in an easy cellular setting. This biosensor is suitable for high-throughput applications and a valuable resource in the researcher's toolkit.7 p.application/pdfengcc by (c) Morató Arús, Xavier, et al.; American Chemical Society, 2023https://creativecommons.org/licenses/by/4.0/Proteïnes portadoresDisseny de medicamentsBiosensorsOligòmersCarrier proteinsDrug designBiosensorsOligomersinfo:eu-repo/semantics/article7364632023-07-14info:eu-repo/semantics/openAccess37191585