title: Functional selectivity of allosteric interactions within G protein-coupled receptor oligomers: the dopamine D1-D3 receptor heterotetramer.
abstract: The dopamine D1 receptor-D3 receptor (D1R-D3R) heteromer is being considered as a potential therapeutic target for neuropsychiatric disorders. Previous studies suggested that this heteromer could be involved in the ability of D3R agonists to potentiate locomotor activation induced by D1R agonists. It has also been postulated that its overexpression plays a role in L-dopa-induced dyskinesia and in drug addiction. However, little is known about its biochemical properties. By combining bioluminescence resonance energy transfer, bimolecular complementation techniques, and cell-signaling experiments in transfected cells, evidence was obtained for a tetrameric stoichiometry of the D1R-D3R heteromer, constituted by two interacting D1R and D3R homodimers coupled to Gs and Gi proteins, respectively. Coactivation of both receptors led to the canonical negative interaction at the level of adenylyl cyclase signaling, to a strong recruitment of β-arrestin-1, and to a positive cross talk of D1R and D3R agonists at the level of mitogen-activated protein kinase (MAPK) signaling. Furthermore, D1R or D3R antagonists counteracted β-arrestin-1 recruitment and MAPK activation induced by D3R and D1R agonists, respectively (cross-antagonism). Positive cross talk and cross-antagonism at the MAPK level were counteracted by specific synthetic peptides with amino acid sequences corresponding to D1R transmembrane (TM) domains TM5 and TM6, which also selectively modified the quaternary structure of the D1R-D3R heteromer, as demonstrated by complementation of hemiproteins of yellow fluorescence protein fused to D1R and D3R. These results demonstrate functional selectivity of allosteric modulations within the D1R-D3R heteromer, which can be involved with the reported behavioral synergism of D1R and D3R agonists.
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Xavier
authors.initials: X
authors.lastname: Guitart
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Gemma
authors.initials: G
authors.lastname: Navarro
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Estefania
authors.initials: E
authors.lastname: Moreno
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Hideaki
authors.initials: H
authors.lastname: Yano
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Ning-Sheng
authors.initials: NS
authors.lastname: Cai
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Marta
authors.initials: M
authors.lastname: Sánchez-Soto
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Sandeep
authors.initials: S
authors.lastname: Kumar-Barodia
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Yamini T
authors.initials: YT
authors.lastname: Naidu
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Josefa
authors.initials: J
authors.lastname: Mallol
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Antoni
authors.initials: A
authors.lastname: Cortés
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Carme
authors.initials: C
authors.lastname: Lluís
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Enric I
authors.initials: EI
authors.lastname: Canela
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Vicent
authors.initials: V
authors.lastname: Casadó
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.).
authors.forename: Peter J
authors.initials: PJ
authors.lastname: McCormick
authors.affiliation: National Institute on Drug Abuse, Intramural Research Program, National Institutes of Health, Department of Health and Human Services, Baltimore, Maryland (X.G., H.Y., N.-S.C., M.S.-S., S.K.-B., Y.T.N., S.F.); Centro de Investigación Biomédica en Red Sobre Enfermedades Neurodegenerativas and Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Barcelona, Barcelona, Spain (G.N., E.M., J.M., A.C., C.L., E.I.C., V.C., P.J.M.); and School of Pharmacy, University of East Anglia, Norwich, United Kingdom (P.J.M.) sferre@intra.nida.nih.gov.
authors.forename: Sergi
authors.initials: S
authors.lastname: Ferré
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D000494:Allosteric Regulation; D000495:Allosteric Site; D019390:Arrestins / Q000378:metabolism; D018491:Dopamine Agonists / Q000494:pharmacology; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D020935:MAP Kinase Signaling System; D011485:Protein Binding; D055503:Protein Multimerization; D017447:Receptors, Dopamine D1 / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D050637:Receptors, Dopamine D3 / Q000819:agonists / Q000737:chemistry / Q000378:metabolism*; D000071559:beta-Arrestin 1; D000071557:beta-Arrestins
chemical_list: C000605253:ARRB1 protein, human; D019390:Arrestins; C583260:DRD1 protein, human; C493593:DRD3 protein, human; D018491:Dopamine Agonists; D017447:Receptors, Dopamine D1; D050637:Receptors, Dopamine D3; D000071559:beta-Arrestin 1; D000071557:beta-Arrestins; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D019205:GTP-Binding Protein alpha Subunits, Gs; D000262:Adenylyl Cyclases
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