Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors

The mechanism of action of cannabidiol (CBD), the main non-psychotropic component of Cannabis sativa L., is not completely understood. First assumed that the compound was acting via cannabinoid CB2 receptors (CB(2)Rs) it is now suggested that it interacts with non-cannabinoid G-protein-coupled recep...

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Autores: Martínez-Pinilla, Eva, Varani, Katia, Reyes Resina, Irene, Angelats Canals, Edgar, Vincenzi, Fabrizio, Ferreiro-Vera, Carlos, Oyarzabal, Julen, Canela Campos, Enric I. (Enric Isidre), 1949-, Lanciego, José Luis, Nadal, Xavier, Navarro Brugal, Gemma, Borea, Pier Andrea, Franco Fernández, Rafael
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2017
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/122632
Acceso en línea:https://hdl.handle.net/2445/122632
Access Level:acceso abierto
Palabra clave:Receptors cel·lulars
Proteïnes G
Cell receptors
G Proteins
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spelling Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 ReceptorsMartínez-Pinilla, EvaVarani, KatiaReyes Resina, IreneAngelats Canals, EdgarVincenzi, FabrizioFerreiro-Vera, CarlosOyarzabal, JulenCanela Campos, Enric I. (Enric Isidre), 1949-Lanciego, José LuisNadal, XavierNavarro Brugal, GemmaBorea, Pier AndreaFranco Fernández, RafaelReceptors cel·lularsProteïnes GCell receptorsG ProteinsThe mechanism of action of cannabidiol (CBD), the main non-psychotropic component of Cannabis sativa L., is not completely understood. First assumed that the compound was acting via cannabinoid CB2 receptors (CB(2)Rs) it is now suggested that it interacts with non-cannabinoid G-protein-coupled receptors (GPCRs); however, CBD does not bind with high affinity to the orthosteric site of any GPCR. To search for alternative explanations, we tested CBD as a potential allosteric ligand of CB2R. Radioligand and non-radioactive homogeneous binding, intracellular cAMP determination and ERK1/2 phosphorylation assays were undertaken in heterologous systems expressing the human version of CB2R. Using membrane preparations from CB2R-expressing HEK-293T (human embryonic kidney 293T) cells, we confirmed that CBD does not bind with high affinity to the orthosteric site of the human CB2R where the synthetic cannabinoid, [H-3]-WIN 55,212-2, binds. CBD was, however, able to produce minor but consistent reduction in the homogeneous binding assays in living cells using the fluorophore-conjugated CB2R-selective compound, CM-157. The effect on binding to CB2R-expressing living cells was different to that exerted by the orthosteric antagonist, SR144528, which decreased the maximum binding without changing the K-D. CBD at nanomolar concentrations was also able to significantly reduce the effect of the selective CB2R agonist, JWH133, on forskolin-induced intracellular cAMP levels and on activation of the MAP kinase pathway. These results may help to understand CBD mode of action and may serve to revisit its therapeutic possibilities.Frontiers Media2018201820172018info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion10 p.application/pdfapplication/pdfhttps://hdl.handle.net/2445/122632Articles publicats en revistes (Bioquímica i Biomedicina Molecular)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésReproducció del document publicat a: https://doi.org/10.3389/fphar.2017.00744Frontiers in Pharmacology, 2017, vol. 8https://doi.org/10.3389/fphar.2017.00744cc-by (c) Martinez-Pinilla, Eva et al., 2017http://creativecommons.org/licenses/by/3.0/esinfo:eu-repo/semantics/openAccessoai:recercat.cat:2445/1226322026-05-29T05:05:01Z
dc.title.none.fl_str_mv Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
title Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
spellingShingle Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
Martínez-Pinilla, Eva
Receptors cel·lulars
Proteïnes G
Cell receptors
G Proteins
title_short Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
title_full Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
title_fullStr Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
title_full_unstemmed Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
title_sort Binding and Signaling Studies Disclose a Potential Allosteric Site for Cannabidiol in Cannabinoid CB2 Receptors
dc.creator.none.fl_str_mv Martínez-Pinilla, Eva
Varani, Katia
Reyes Resina, Irene
Angelats Canals, Edgar
Vincenzi, Fabrizio
Ferreiro-Vera, Carlos
Oyarzabal, Julen
Canela Campos, Enric I. (Enric Isidre), 1949-
Lanciego, José Luis
Nadal, Xavier
Navarro Brugal, Gemma
Borea, Pier Andrea
Franco Fernández, Rafael
author Martínez-Pinilla, Eva
author_facet Martínez-Pinilla, Eva
Varani, Katia
Reyes Resina, Irene
Angelats Canals, Edgar
Vincenzi, Fabrizio
Ferreiro-Vera, Carlos
Oyarzabal, Julen
Canela Campos, Enric I. (Enric Isidre), 1949-
Lanciego, José Luis
Nadal, Xavier
Navarro Brugal, Gemma
Borea, Pier Andrea
Franco Fernández, Rafael
author_role author
author2 Varani, Katia
Reyes Resina, Irene
Angelats Canals, Edgar
Vincenzi, Fabrizio
Ferreiro-Vera, Carlos
Oyarzabal, Julen
Canela Campos, Enric I. (Enric Isidre), 1949-
Lanciego, José Luis
Nadal, Xavier
Navarro Brugal, Gemma
Borea, Pier Andrea
Franco Fernández, Rafael
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Receptors cel·lulars
Proteïnes G
Cell receptors
G Proteins
topic Receptors cel·lulars
Proteïnes G
Cell receptors
G Proteins
description The mechanism of action of cannabidiol (CBD), the main non-psychotropic component of Cannabis sativa L., is not completely understood. First assumed that the compound was acting via cannabinoid CB2 receptors (CB(2)Rs) it is now suggested that it interacts with non-cannabinoid G-protein-coupled receptors (GPCRs); however, CBD does not bind with high affinity to the orthosteric site of any GPCR. To search for alternative explanations, we tested CBD as a potential allosteric ligand of CB2R. Radioligand and non-radioactive homogeneous binding, intracellular cAMP determination and ERK1/2 phosphorylation assays were undertaken in heterologous systems expressing the human version of CB2R. Using membrane preparations from CB2R-expressing HEK-293T (human embryonic kidney 293T) cells, we confirmed that CBD does not bind with high affinity to the orthosteric site of the human CB2R where the synthetic cannabinoid, [H-3]-WIN 55,212-2, binds. CBD was, however, able to produce minor but consistent reduction in the homogeneous binding assays in living cells using the fluorophore-conjugated CB2R-selective compound, CM-157. The effect on binding to CB2R-expressing living cells was different to that exerted by the orthosteric antagonist, SR144528, which decreased the maximum binding without changing the K-D. CBD at nanomolar concentrations was also able to significantly reduce the effect of the selective CB2R agonist, JWH133, on forskolin-induced intracellular cAMP levels and on activation of the MAP kinase pathway. These results may help to understand CBD mode of action and may serve to revisit its therapeutic possibilities.
publishDate 2017
dc.date.none.fl_str_mv 2017
2018
2018
2018
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/122632
url https://hdl.handle.net/2445/122632
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.3389/fphar.2017.00744
Frontiers in Pharmacology, 2017, vol. 8
https://doi.org/10.3389/fphar.2017.00744
dc.rights.none.fl_str_mv cc-by (c) Martinez-Pinilla, Eva et al., 2017
http://creativecommons.org/licenses/by/3.0/es
info:eu-repo/semantics/openAccess
rights_invalid_str_mv cc-by (c) Martinez-Pinilla, Eva et al., 2017
http://creativecommons.org/licenses/by/3.0/es
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 10 p.
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv Articles publicats en revistes (Bioquímica i Biomedicina Molecular)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.81155

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