2026-04-14T02:43:20Zhttps://recercat.cat/oai/request
oai:recercat.cat:2445/1919752025-12-04T21:12:14Zcom_2072_1057col_2072_478781col_2072_478917
RECERCAT author Casajuana-Martin, Nil author Navarro Brugal, Gemma author González Ureña, A. author Llinàs Del Torrent Masachs, Clàudia author Gómez-Autet, Marc author Quintana García, Aleix author Franco Fernández, Rafael author Pardo, Leonardo 2023-01-10T08:44:44Z2023-11-28T06:10:21Z2022-11-282023-01-10T08:44:44Z 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. (c) American Chemical Society , 2022 http://creativecommons.org/licenses/by/3.0/es/ info:eu-repo/semantics/openAccess Dinàmica molecular Bicapes lipídiques Mutació (Biologia) Molecular dynamics Lipid bilayers Mutation (Biology) A Single Point Mutation Blocks the Entrance of Ligands to the Cannabinoid CB2 Receptor via the Lipid Bilayer info:eu-repo/semantics/article info:eu-repo/semantics/acceptedVersion