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oai:recercat.cat:2445/1919752025-12-04T21:12:14Zcom_2072_1057col_2072_478781col_2072_478917

A Single Point Mutation Blocks the Entrance of Ligands to the Cannabinoid CB2 Receptor via the Lipid Bilayer Casajuana-Martin, Nil Navarro Brugal, Gemma González Ureña, A. Llinàs Del Torrent Masachs, Clàudia Gómez-Autet, Marc Quintana García, Aleix Franco Fernández, Rafael Pardo, Leonardo Dinàmica molecular Bicapes lipídiques Mutació (Biologia) Molecular dynamics Lipid bilayers Mutation (Biology) Molecular dynamic (MD) simulations have become a common tool to study the pathway of ligand entry to the orthosteric binding site of G protein-coupled receptors. Here, we have combined MD simulations and site-directed mutagenesis to study the binding process of the potent JWH-133 agonist to the cannabinoid CB2 receptor (CB2R). In CB2R, the N-terminus and extracellular loop 2 fold over the ligand binding pocket, blocking access to the binding cavity from the extracellular environment. We, thus, hypothesized that the binding pathway is a multistage process consisting of the hydrophobic ligand diffusing in the lipid bilayer to contact a lipid-facing vestibule, from which the ligand enters an allosteric site inside the transmembrane bundle through a tunnel formed between TMs 1 and 7 and finally moving from the allosteric to the orthosteric binding cavity. This pathway was experimentally validated by the Ala2827.36Phe mutation that blocks the entrance of the ligand, as JWH-133 was not able to decrease the forskolin-induced cAMP levels in cells expressing the mutant receptor. This proposed ligand entry pathway defines transient binding sites that are potential cavities for the design of synthetic modulators. 2023-01-10T08:44:44Z 2023-11-28T06:10:21Z 2022-11-28 2023-01-10T08:44:44Z info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion Versió postprint del document publicat a: https://doi.org/10.1021/acs.jcim.2c00865 Journal of Chemical Information and Modeling, 2022, vol. 62, num. 22, p. 5771-5779 https://doi.org/10.1021/acs.jcim.2c00865 (c) American Chemical Society , 2022 http://creativecommons.org/licenses/by/3.0/es/ info:eu-repo/semantics/openAccess American Chemical Society Articles publicats en revistes (Bioquímica i Biomedicina Molecular)