Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer

There is evidence for strong functional antagonistic interactions between adenosine A2A receptors (A2ARs) and dopamine D2 receptors (D2Rs). Although a close physical interaction between both receptors has recently been shown using co-immunoprecipitation and co-localization assays, the existence of a...

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Autores: Canals Buj, Meritxell, Marcellino, Daniel, Fanelli, Francesca, Ciruela Alférez, Francisco, Benedetti, Piero de, Goldberg, Steven R., Neve, Kim, Fuxe, Kjell, Agnati, Luigi F., Woods, Amina S., Ferré, Sergi, Lluís i Biset, Carme, Bouvier, Michael, Franco Fernández, Rafael
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2003
País:España
Recursos: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:2445/176963
Acesso em linha:https://hdl.handle.net/2445/176963
Access Level:acceso abierto
Palavra-chave:Transferència d'energia
Adenosina
Dopamina
Metabolisme
Energy transfer
Adenosine
Dopamine
Metabolism
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spelling Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transferCanals Buj, MeritxellMarcellino, DanielFanelli, FrancescaCiruela Alférez, FranciscoBenedetti, Piero deGoldberg, Steven R.Neve, KimFuxe, KjellAgnati, Luigi F.Woods, Amina S.Ferré, SergiLluís i Biset, CarmeBouvier, MichaelFranco Fernández, RafaelTransferència d'energiaAdenosinaDopaminaMetabolismeEnergy transferAdenosineDopamineMetabolismThere is evidence for strong functional antagonistic interactions between adenosine A2A receptors (A2ARs) and dopamine D2 receptors (D2Rs). Although a close physical interaction between both receptors has recently been shown using co-immunoprecipitation and co-localization assays, the existence of a A2AR-D2R protein-protein interaction still had to be demonstrated in intact living cells. In the present work, fluorescence resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET) techniques were used to confirm the occurrence of A2AR-D2R interactions in co-transfected cells. The degree of A2AR-D2R heteromerization, measured by BRET, did not vary after receptor activation with selective agonists, alone or in combination. BRET competition experiments were performed using a chimeric D2R-D1R in which helices 5 and 6, the third intracellular loop (I3), and the third extracellular loop (E3) of the D2R were replaced by those of the dopamine D1 receptor (D1R). Although the wild type D2R was able to decrease the BRET signal, the chimera failed to achieve any effect. This suggests that the helix 5-I3-helix 6-E3 portion of D2R holds the site(s) for interaction with A2AR. Modeling of A2AR and D2R using a modified rhodopsin template followed by molecular dynamics and docking simulations gave essentially two different possible modes of interaction between D2R and A2AR. In the most probable one, helix 5 and/or helix 6 and the N-terminal portion of I3 from D2R approached helix 4 and the C-terminal portion of the C-tail from the A2AR, respectively.American Society for Biochemistry and Molecular Biology2021202120032021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion9 p.application/pdfapplication/pdfhttps://hdl.handle.net/2445/176963Articles publicats en revistes (Patologia i Terapèutica Experimental)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/10.1074/jbc.M306451200Journal of Biological Chemistry, 2003, vol. 278, num. 47, p. 46741-46749https://doi.org/10.1074/jbc.M306451200(c) American Society for Biochemistry and Molecular Biology, 2003info:eu-repo/semantics/openAccessoai:recercat.cat:2445/1769632026-05-29T05:05:01Z
dc.title.none.fl_str_mv Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
title Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
spellingShingle Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
Canals Buj, Meritxell
Transferència d'energia
Adenosina
Dopamina
Metabolisme
Energy transfer
Adenosine
Dopamine
Metabolism
title_short Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
title_full Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
title_fullStr Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
title_full_unstemmed Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
title_sort Adenosine A2A-dopamine D2 receptor-receptor heteromerization: qualitative and quantitative assessment by fluorescence and bioluminescence energy transfer
dc.creator.none.fl_str_mv Canals Buj, Meritxell
Marcellino, Daniel
Fanelli, Francesca
Ciruela Alférez, Francisco
Benedetti, Piero de
Goldberg, Steven R.
Neve, Kim
Fuxe, Kjell
Agnati, Luigi F.
Woods, Amina S.
Ferré, Sergi
Lluís i Biset, Carme
Bouvier, Michael
Franco Fernández, Rafael
author Canals Buj, Meritxell
author_facet Canals Buj, Meritxell
Marcellino, Daniel
Fanelli, Francesca
Ciruela Alférez, Francisco
Benedetti, Piero de
Goldberg, Steven R.
Neve, Kim
Fuxe, Kjell
Agnati, Luigi F.
Woods, Amina S.
Ferré, Sergi
Lluís i Biset, Carme
Bouvier, Michael
Franco Fernández, Rafael
author_role author
author2 Marcellino, Daniel
Fanelli, Francesca
Ciruela Alférez, Francisco
Benedetti, Piero de
Goldberg, Steven R.
Neve, Kim
Fuxe, Kjell
Agnati, Luigi F.
Woods, Amina S.
Ferré, Sergi
Lluís i Biset, Carme
Bouvier, Michael
Franco Fernández, Rafael
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Transferència d'energia
Adenosina
Dopamina
Metabolisme
Energy transfer
Adenosine
Dopamine
Metabolism
topic Transferència d'energia
Adenosina
Dopamina
Metabolisme
Energy transfer
Adenosine
Dopamine
Metabolism
description There is evidence for strong functional antagonistic interactions between adenosine A2A receptors (A2ARs) and dopamine D2 receptors (D2Rs). Although a close physical interaction between both receptors has recently been shown using co-immunoprecipitation and co-localization assays, the existence of a A2AR-D2R protein-protein interaction still had to be demonstrated in intact living cells. In the present work, fluorescence resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET) techniques were used to confirm the occurrence of A2AR-D2R interactions in co-transfected cells. The degree of A2AR-D2R heteromerization, measured by BRET, did not vary after receptor activation with selective agonists, alone or in combination. BRET competition experiments were performed using a chimeric D2R-D1R in which helices 5 and 6, the third intracellular loop (I3), and the third extracellular loop (E3) of the D2R were replaced by those of the dopamine D1 receptor (D1R). Although the wild type D2R was able to decrease the BRET signal, the chimera failed to achieve any effect. This suggests that the helix 5-I3-helix 6-E3 portion of D2R holds the site(s) for interaction with A2AR. Modeling of A2AR and D2R using a modified rhodopsin template followed by molecular dynamics and docking simulations gave essentially two different possible modes of interaction between D2R and A2AR. In the most probable one, helix 5 and/or helix 6 and the N-terminal portion of I3 from D2R approached helix 4 and the C-terminal portion of the C-tail from the A2AR, respectively.
publishDate 2003
dc.date.none.fl_str_mv 2003
2021
2021
2021
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/2445/176963
url https://hdl.handle.net/2445/176963
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/10.1074/jbc.M306451200
Journal of Biological Chemistry, 2003, vol. 278, num. 47, p. 46741-46749
https://doi.org/10.1074/jbc.M306451200
dc.rights.none.fl_str_mv (c) American Society for Biochemistry and Molecular Biology, 2003
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) American Society for Biochemistry and Molecular Biology, 2003
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 9 p.
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv Articles publicats en revistes (Patologia i Terapèutica Experimental)
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 15,811543

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