A Single Point Mutation Blocks the Entrance of Ligands to the Cannabinoid CB Receptor via the Lipid Bilayer
Casajuana-Martin, Nil (Universitat Autònoma de Barcelona)
Navarro Brugal, Gemma (Instituto de Salud Carlos III)
Gonzalez, Angel (Universitat Autònoma de Barcelona)
Llinas del Torrent, Claudia (Universitat Autònoma de Barcelona)
Gómez-Autet, Marc (Universitat Autònoma de Barcelona)
Quintana García, Aleix (Universitat Autònoma de Barcelona)
Franco, Rafael (Instituto de Salud Carlos III)
Pardo Carrasco, Leonardo (Universitat Autònoma de Barcelona)

Date:	2022
Abstract:	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 CB receptor (CBR). In CBR, 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 Ala282 7. 36 Phe 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.
Grants:	Ministerio de Economía y Competitividad BES-2017-081872 Agencia Estatal de Investigación RTI2018-094204-B-I00 Agencia Estatal de Investigación PID2020-113430RB-I00 Agencia Estatal de Investigación PID2019-109240RB-I00
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Published in:	Journal of Chemical Information and Modeling, Vol. 62 (october 2022) , p. 5771-5779, ISSN 1549-960X

DOI: 10.1021/acs.jcim.2c00865
PMID: 36302505

9 p, 5.0 MB

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