Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors

Autores
Pleli, Thomas; Mondorf, Antonia; Ferreiros, Nerea; Thomas, Dominique; Dvorak, Karel; Biondi, Ricardo Miguel; Heringdorf, Dagmar Meyer zu; Zeuzem, Stefan; Geisslinger, Gerd; Zimmermann, Herbert; Waidmann, Oliver; Piiper, Albrecht
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
AbstractBACKGROUND/AIMS:Signaling of Gs protein-coupled receptors (GsPCRs) is accomplished by stimulation of adenylyl cyclase, causing an increase of the intracellular cAMP concentration, activation of the intracellular cAMP effectors protein kinase A (PKA) and Epac, and an efflux of cAMP, the function of which is still unclear.METHODS:Activation of adenylyl cyclase by GsPCR agonists or cholera toxin was monitored by measurement of the intracellular cAMP concentration by ELISA, anti-phospho-PKA substrate motif phosphorylation by immunoblotting, and an Epac-FRET assay in the presence and absence of adenosine receptor antagonists or ecto-nucleotide phosphodiesterase/pyrophosphatase2 (eNPP2) inhibitors. The production of AMP from cAMP by recombinant eNPP2 was measured by HPLC. Extracellular adenosine was determined by LC-MS/MS, extracellular ATP by luciferase and LC-MS/MS. The expression of eNPP isoenzymes 1-3 was examined by RT-PCR. The expression of multidrug resistance protein 4 was suppressed by siRNA.RESULTS:Here we show that the activation of GsPCRs and the GsPCRs-independent activation of Gs proteins and adenylyl cyclase by cholera toxin induce stimulation of cell surface adenosine receptors (A2A or A2B adenosine receptors). In PC12 cells stimulation of adenylyl cyclase by GsPCR or cholera toxin caused activation of A2A adenosine receptors by an autocrine signaling pathway involving cAMP efflux through multidrug resistance protein 4 and hydrolysis of released cAMP to AMP by eNPP2. In contrast, in PC3 cells cholera toxin- and GsPCR-induced stimulation of adenylyl cyclase resulted in the activation of A2B adenosine receptors.CONCLUSION:Our findings show that stimulation of adenylyl cyclase causes a remarkable activation of cell surface adenosine receptors.
Fil: Pleli, Thomas. Goethe Universitat Frankfurt; Alemania
Fil: Mondorf, Antonia. Goethe Universitat Frankfurt; Alemania
Fil: Ferreiros, Nerea. Goethe Universitat Frankfurt; Alemania
Fil: Thomas, Dominique. Goethe Universitat Frankfurt; Alemania
Fil: Dvorak, Karel. Goethe Universitat Frankfurt; Alemania
Fil: Biondi, Ricardo Miguel. Goethe Universitat Frankfurt; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Heringdorf, Dagmar Meyer zu. Goethe Universitat Frankfurt; Alemania
Fil: Zeuzem, Stefan. Goethe Universitat Frankfurt; Alemania
Fil: Geisslinger, Gerd. Goethe Universitat Frankfurt; Alemania
Fil: Zimmermann, Herbert. Goethe Universitat Frankfurt; Alemania
Fil: Waidmann, Oliver. Goethe Universitat Frankfurt; Alemania
Fil: Piiper, Albrecht. Goethe Universitat Frankfurt; Alemania
Materia
Adenosine
Adenylyl Cyclase
cAMP
autocrine
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/88238
Acceder
id CONICETDig_1cd4087793c0a1b70329a89fb7cb68ed
oai_identifier_str oai:ri.conicet.gov.ar:11336/88238
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Activation of Adenylyl Cyclase Causes Stimulation of Adenosine ReceptorsPleli, ThomasMondorf, AntoniaFerreiros, NereaThomas, DominiqueDvorak, KarelBiondi, Ricardo MiguelHeringdorf, Dagmar Meyer zuZeuzem, StefanGeisslinger, GerdZimmermann, HerbertWaidmann, OliverPiiper, AlbrechtAdenosineAdenylyl CyclasecAMPautocrinehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1AbstractBACKGROUND/AIMS:Signaling of Gs protein-coupled receptors (GsPCRs) is accomplished by stimulation of adenylyl cyclase, causing an increase of the intracellular cAMP concentration, activation of the intracellular cAMP effectors protein kinase A (PKA) and Epac, and an efflux of cAMP, the function of which is still unclear.METHODS:Activation of adenylyl cyclase by GsPCR agonists or cholera toxin was monitored by measurement of the intracellular cAMP concentration by ELISA, anti-phospho-PKA substrate motif phosphorylation by immunoblotting, and an Epac-FRET assay in the presence and absence of adenosine receptor antagonists or ecto-nucleotide phosphodiesterase/pyrophosphatase2 (eNPP2) inhibitors. The production of AMP from cAMP by recombinant eNPP2 was measured by HPLC. Extracellular adenosine was determined by LC-MS/MS, extracellular ATP by luciferase and LC-MS/MS. The expression of eNPP isoenzymes 1-3 was examined by RT-PCR. The expression of multidrug resistance protein 4 was suppressed by siRNA.RESULTS:Here we show that the activation of GsPCRs and the GsPCRs-independent activation of Gs proteins and adenylyl cyclase by cholera toxin induce stimulation of cell surface adenosine receptors (A2A or A2B adenosine receptors). In PC12 cells stimulation of adenylyl cyclase by GsPCR or cholera toxin caused activation of A2A adenosine receptors by an autocrine signaling pathway involving cAMP efflux through multidrug resistance protein 4 and hydrolysis of released cAMP to AMP by eNPP2. In contrast, in PC3 cells cholera toxin- and GsPCR-induced stimulation of adenylyl cyclase resulted in the activation of A2B adenosine receptors.CONCLUSION:Our findings show that stimulation of adenylyl cyclase causes a remarkable activation of cell surface adenosine receptors.Fil: Pleli, Thomas. Goethe Universitat Frankfurt; AlemaniaFil: Mondorf, Antonia. Goethe Universitat Frankfurt; AlemaniaFil: Ferreiros, Nerea. Goethe Universitat Frankfurt; AlemaniaFil: Thomas, Dominique. Goethe Universitat Frankfurt; AlemaniaFil: Dvorak, Karel. Goethe Universitat Frankfurt; AlemaniaFil: Biondi, Ricardo Miguel. Goethe Universitat Frankfurt; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Heringdorf, Dagmar Meyer zu. Goethe Universitat Frankfurt; AlemaniaFil: Zeuzem, Stefan. Goethe Universitat Frankfurt; AlemaniaFil: Geisslinger, Gerd. Goethe Universitat Frankfurt; AlemaniaFil: Zimmermann, Herbert. Goethe Universitat Frankfurt; AlemaniaFil: Waidmann, Oliver. Goethe Universitat Frankfurt; AlemaniaFil: Piiper, Albrecht. Goethe Universitat Frankfurt; AlemaniaKarger2018-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/88238Pleli, Thomas; Mondorf, Antonia; Ferreiros, Nerea; Thomas, Dominique; Dvorak, Karel; et al.; Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors; Karger; Cellular Physiology and Biochemistry; 45; 6; 5-2018; 2516-25281015-8987CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.karger.com/Article/FullText/488270info:eu-repo/semantics/altIdentifier/doi/10.1159/000488270info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:39:46Zoai:ri.conicet.gov.ar:11336/88238instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:39:47.067CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
title Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
spellingShingle Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
Pleli, Thomas
Adenosine
Adenylyl Cyclase
cAMP
autocrine
title_short Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
title_full Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
title_fullStr Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
title_full_unstemmed Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
title_sort Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
dc.creator.none.fl_str_mv Pleli, Thomas
Mondorf, Antonia
Ferreiros, Nerea
Thomas, Dominique
Dvorak, Karel
Biondi, Ricardo Miguel
Heringdorf, Dagmar Meyer zu
Zeuzem, Stefan
Geisslinger, Gerd
Zimmermann, Herbert
Waidmann, Oliver
Piiper, Albrecht
author Pleli, Thomas
author_facet Pleli, Thomas
Mondorf, Antonia
Ferreiros, Nerea
Thomas, Dominique
Dvorak, Karel
Biondi, Ricardo Miguel
Heringdorf, Dagmar Meyer zu
Zeuzem, Stefan
Geisslinger, Gerd
Zimmermann, Herbert
Waidmann, Oliver
Piiper, Albrecht
author_role author
author2 Mondorf, Antonia
Ferreiros, Nerea
Thomas, Dominique
Dvorak, Karel
Biondi, Ricardo Miguel
Heringdorf, Dagmar Meyer zu
Zeuzem, Stefan
Geisslinger, Gerd
Zimmermann, Herbert
Waidmann, Oliver
Piiper, Albrecht
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Adenosine
Adenylyl Cyclase
cAMP
autocrine
topic Adenosine
Adenylyl Cyclase
cAMP
autocrine
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv AbstractBACKGROUND/AIMS:Signaling of Gs protein-coupled receptors (GsPCRs) is accomplished by stimulation of adenylyl cyclase, causing an increase of the intracellular cAMP concentration, activation of the intracellular cAMP effectors protein kinase A (PKA) and Epac, and an efflux of cAMP, the function of which is still unclear.METHODS:Activation of adenylyl cyclase by GsPCR agonists or cholera toxin was monitored by measurement of the intracellular cAMP concentration by ELISA, anti-phospho-PKA substrate motif phosphorylation by immunoblotting, and an Epac-FRET assay in the presence and absence of adenosine receptor antagonists or ecto-nucleotide phosphodiesterase/pyrophosphatase2 (eNPP2) inhibitors. The production of AMP from cAMP by recombinant eNPP2 was measured by HPLC. Extracellular adenosine was determined by LC-MS/MS, extracellular ATP by luciferase and LC-MS/MS. The expression of eNPP isoenzymes 1-3 was examined by RT-PCR. The expression of multidrug resistance protein 4 was suppressed by siRNA.RESULTS:Here we show that the activation of GsPCRs and the GsPCRs-independent activation of Gs proteins and adenylyl cyclase by cholera toxin induce stimulation of cell surface adenosine receptors (A2A or A2B adenosine receptors). In PC12 cells stimulation of adenylyl cyclase by GsPCR or cholera toxin caused activation of A2A adenosine receptors by an autocrine signaling pathway involving cAMP efflux through multidrug resistance protein 4 and hydrolysis of released cAMP to AMP by eNPP2. In contrast, in PC3 cells cholera toxin- and GsPCR-induced stimulation of adenylyl cyclase resulted in the activation of A2B adenosine receptors.CONCLUSION:Our findings show that stimulation of adenylyl cyclase causes a remarkable activation of cell surface adenosine receptors.
Fil: Pleli, Thomas. Goethe Universitat Frankfurt; Alemania
Fil: Mondorf, Antonia. Goethe Universitat Frankfurt; Alemania
Fil: Ferreiros, Nerea. Goethe Universitat Frankfurt; Alemania
Fil: Thomas, Dominique. Goethe Universitat Frankfurt; Alemania
Fil: Dvorak, Karel. Goethe Universitat Frankfurt; Alemania
Fil: Biondi, Ricardo Miguel. Goethe Universitat Frankfurt; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina
Fil: Heringdorf, Dagmar Meyer zu. Goethe Universitat Frankfurt; Alemania
Fil: Zeuzem, Stefan. Goethe Universitat Frankfurt; Alemania
Fil: Geisslinger, Gerd. Goethe Universitat Frankfurt; Alemania
Fil: Zimmermann, Herbert. Goethe Universitat Frankfurt; Alemania
Fil: Waidmann, Oliver. Goethe Universitat Frankfurt; Alemania
Fil: Piiper, Albrecht. Goethe Universitat Frankfurt; Alemania
description AbstractBACKGROUND/AIMS:Signaling of Gs protein-coupled receptors (GsPCRs) is accomplished by stimulation of adenylyl cyclase, causing an increase of the intracellular cAMP concentration, activation of the intracellular cAMP effectors protein kinase A (PKA) and Epac, and an efflux of cAMP, the function of which is still unclear.METHODS:Activation of adenylyl cyclase by GsPCR agonists or cholera toxin was monitored by measurement of the intracellular cAMP concentration by ELISA, anti-phospho-PKA substrate motif phosphorylation by immunoblotting, and an Epac-FRET assay in the presence and absence of adenosine receptor antagonists or ecto-nucleotide phosphodiesterase/pyrophosphatase2 (eNPP2) inhibitors. The production of AMP from cAMP by recombinant eNPP2 was measured by HPLC. Extracellular adenosine was determined by LC-MS/MS, extracellular ATP by luciferase and LC-MS/MS. The expression of eNPP isoenzymes 1-3 was examined by RT-PCR. The expression of multidrug resistance protein 4 was suppressed by siRNA.RESULTS:Here we show that the activation of GsPCRs and the GsPCRs-independent activation of Gs proteins and adenylyl cyclase by cholera toxin induce stimulation of cell surface adenosine receptors (A2A or A2B adenosine receptors). In PC12 cells stimulation of adenylyl cyclase by GsPCR or cholera toxin caused activation of A2A adenosine receptors by an autocrine signaling pathway involving cAMP efflux through multidrug resistance protein 4 and hydrolysis of released cAMP to AMP by eNPP2. In contrast, in PC3 cells cholera toxin- and GsPCR-induced stimulation of adenylyl cyclase resulted in the activation of A2B adenosine receptors.CONCLUSION:Our findings show that stimulation of adenylyl cyclase causes a remarkable activation of cell surface adenosine receptors.
publishDate 2018
dc.date.none.fl_str_mv 2018-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/88238
Pleli, Thomas; Mondorf, Antonia; Ferreiros, Nerea; Thomas, Dominique; Dvorak, Karel; et al.; Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors; Karger; Cellular Physiology and Biochemistry; 45; 6; 5-2018; 2516-2528
1015-8987
CONICET Digital
CONICET
url http://hdl.handle.net/11336/88238
identifier_str_mv Pleli, Thomas; Mondorf, Antonia; Ferreiros, Nerea; Thomas, Dominique; Dvorak, Karel; et al.; Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors; Karger; Cellular Physiology and Biochemistry; 45; 6; 5-2018; 2516-2528
1015-8987
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.karger.com/Article/FullText/488270
info:eu-repo/semantics/altIdentifier/doi/10.1159/000488270
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Karger
publisher.none.fl_str_mv Karger
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1846082885260410880
score 13.22299

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