TY - JOUR
T1 - Allosterism in the adenosine A2A and cannabinoid CB2 heteromer
AU - Llinas del Torrent, Claudia
AU - Raïch, Iu
AU - Gonzalez, Angel
AU - Lillo, Jaume
AU - Casajuana-Martin, Nil
AU - Franco, Rafael
AU - Pardo, Leonardo
AU - Navarro, Gemma
N1 - Publisher Copyright:
© 2024 The Author(s). British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.
PY - 2024/7/23
Y1 - 2024/7/23
N2 - Background and Purpose: Allosterism is a regulatory mechanism for GPCRs that can be attained by ligand-binding or protein–protein interactions with another GPCR. We have studied the influence of the dimer interface on the allosteric properties of the A2A receptor and CB2 receptor heteromer. Experimental Approach: We have evaluated cAMP production, phosphorylation of signal-regulated kinases (pERK1/2), label-free dynamic mass redistribution, β-arrestin 2 recruitment and bimolecular fluorescence complementation assays in the absence and presence of synthetic peptides that disrupt the formation of the heteromer. Molecular dynamic simulations provided converging evidence that the heteromeric interface influences the allosteric properties of the A2AR–CB2R heteromer. Key Results: Apo A2AR blocks agonist-induced signalling of CB2R. The disruptive peptides, with the amino acid sequence of transmembrane (TM) 6 of A2AR or CB2R, facilitate CB2R activation, suggesting that A2AR allosterically prevents the outward movement of TM 6 of CB2R for G protein binding. Significantly, binding of the selective antagonist SCH 58261 to A2AR also facilitated agonist-induced activation of CB2R. Conclusions and Implications: It is proposed that the A2AR–CB2R heteromer contains distinct dimerization interfaces that govern its functional properties. The molecular interface between protomers of the A2AR–CB2R heteromer interconverted from TM 6 for apo or agonist-bound A2AR, blocking CB2R activation, to mainly the TM 1/7 interface for antagonist-bound A2AR, facilitating the independent opening of intracellular cavities for G protein binding. These novel results shed light on a different type of allosteric mechanism and extend the repertoire of GPCR heteromer signalling.
AB - Background and Purpose: Allosterism is a regulatory mechanism for GPCRs that can be attained by ligand-binding or protein–protein interactions with another GPCR. We have studied the influence of the dimer interface on the allosteric properties of the A2A receptor and CB2 receptor heteromer. Experimental Approach: We have evaluated cAMP production, phosphorylation of signal-regulated kinases (pERK1/2), label-free dynamic mass redistribution, β-arrestin 2 recruitment and bimolecular fluorescence complementation assays in the absence and presence of synthetic peptides that disrupt the formation of the heteromer. Molecular dynamic simulations provided converging evidence that the heteromeric interface influences the allosteric properties of the A2AR–CB2R heteromer. Key Results: Apo A2AR blocks agonist-induced signalling of CB2R. The disruptive peptides, with the amino acid sequence of transmembrane (TM) 6 of A2AR or CB2R, facilitate CB2R activation, suggesting that A2AR allosterically prevents the outward movement of TM 6 of CB2R for G protein binding. Significantly, binding of the selective antagonist SCH 58261 to A2AR also facilitated agonist-induced activation of CB2R. Conclusions and Implications: It is proposed that the A2AR–CB2R heteromer contains distinct dimerization interfaces that govern its functional properties. The molecular interface between protomers of the A2AR–CB2R heteromer interconverted from TM 6 for apo or agonist-bound A2AR, blocking CB2R activation, to mainly the TM 1/7 interface for antagonist-bound A2AR, facilitating the independent opening of intracellular cavities for G protein binding. These novel results shed light on a different type of allosteric mechanism and extend the repertoire of GPCR heteromer signalling.
KW - A(2A)R-CB2R heteromer
KW - Allosterism
KW - Modulation
KW - Molecular interface
UR - http://www.scopus.com/inward/record.url?scp=85195100048&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/6847f035-bf03-312f-a8ec-8f8bbc40fbe5/
UR - https://portalrecerca.uab.cat/en/publications/00222672-36c1-4fdd-9bed-345c21a7a1a1
U2 - 10.1111/bph.16502
DO - 10.1111/bph.16502
M3 - Article
C2 - 39044481
AN - SCOPUS:85195100048
SN - 0007-1188
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
ER -