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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/88238
Ver los metadatos del registro completo
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oai:ri.conicet.gov.ar:11336/88238 |
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CONICET Digital (CONICET) |
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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 Técnicas2025-09-29T09:46:11Zoai: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:34982025-09-29 09:46:12.015CONICET 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 |
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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 |
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13.070432 |