The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function

Autores
Cantero, Maria del Rocio; Velázquez, Irina Florencia; Streets, Andrew J.; Ong, Albert C. M.; Cantiello, Horacio Fabio
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Polycystin-2 (PC2) is aTRP-type, Ca2+-permeable non-selective cation channel that plays an important role in Ca2+ signaling in renal and non-renal cells. The effect(s) of the cAMP pathway and kinase mediated phosphorylation of PC2 seem to be relevant to PC2 trafficking and its interaction with polycystin-1. However, the role of PC2 phosphorylation in channel function is still poorly defined. Here we reconstituted apical membranes of term human syncytiotrophoblast (hST), containing endogenous PC2 (PC2hst), and in vitro translated channel protein (PC2iv). Addition of the catalytic subunit of PKA increased by 566% the spontaneous PC2hst channel activity in the presence of ATP. Interestingly, 8-Br-cAMP also stimulated spontaneous PC2hst channel activity in the absence of the exogenous kinase. Either stimulation was inhibited by addition of alkaline phosphatase, which in turn, was reversed by the phosphatase inhibitor vanadate. Neither maneuver modified the single channel conductance but instead increased channel mean open time. PKA directly phosphorylated PC2, which increased the mean open time but not the single channel conductance of the channel. PKA phosphorylation did not modify either R742X truncated or S829A-mutant PC2iv channel function. The data indicate that the cAMP pathway regulates PC2-mediated cation transport in the hST. The relevant PKA site for PC2 channel regulation centers on a single residue serine 829, in the carboxyl terminus.
Fil: Cantero, Maria del Rocio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Velázquez, Irina Florencia. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Streets, Andrew J.. University of Sheffield Medical School; Reino Unido
Fil: Ong, Albert C. M.. University of Sheffield Medical School; Reino Unido
Fil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina
Materia
Cyclic Amp (Camp)
Electrophysiology
Placenta
Protein Kinase a (Pka)
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/38777
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/38777
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel FunctionCantero, Maria del RocioVelázquez, Irina FlorenciaStreets, Andrew J.Ong, Albert C. M.Cantiello, Horacio FabioCyclic Amp (Camp)ElectrophysiologyPlacentaProtein Kinase a (Pka)https://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Polycystin-2 (PC2) is aTRP-type, Ca2+-permeable non-selective cation channel that plays an important role in Ca2+ signaling in renal and non-renal cells. The effect(s) of the cAMP pathway and kinase mediated phosphorylation of PC2 seem to be relevant to PC2 trafficking and its interaction with polycystin-1. However, the role of PC2 phosphorylation in channel function is still poorly defined. Here we reconstituted apical membranes of term human syncytiotrophoblast (hST), containing endogenous PC2 (PC2hst), and in vitro translated channel protein (PC2iv). Addition of the catalytic subunit of PKA increased by 566% the spontaneous PC2hst channel activity in the presence of ATP. Interestingly, 8-Br-cAMP also stimulated spontaneous PC2hst channel activity in the absence of the exogenous kinase. Either stimulation was inhibited by addition of alkaline phosphatase, which in turn, was reversed by the phosphatase inhibitor vanadate. Neither maneuver modified the single channel conductance but instead increased channel mean open time. PKA directly phosphorylated PC2, which increased the mean open time but not the single channel conductance of the channel. PKA phosphorylation did not modify either R742X truncated or S829A-mutant PC2iv channel function. The data indicate that the cAMP pathway regulates PC2-mediated cation transport in the hST. The relevant PKA site for PC2 channel regulation centers on a single residue serine 829, in the carboxyl terminus.Fil: Cantero, Maria del Rocio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Velázquez, Irina Florencia. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Streets, Andrew J.. University of Sheffield Medical School; Reino UnidoFil: Ong, Albert C. M.. University of Sheffield Medical School; Reino UnidoFil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; ArgentinaAmerican Society for Biochemistry and Molecular Biology2015-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/38777Cantero, Maria del Rocio; Velázquez, Irina Florencia; Streets, Andrew J.; Ong, Albert C. M.; Cantiello, Horacio Fabio; The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 290; 39; 9-2015; 23888-238960021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M115.661082info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/290/39/23888.longinfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4583051/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:49:07Zoai:ri.conicet.gov.ar:11336/38777instacron: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:34982025-09-03 09:49:07.529CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
title The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
spellingShingle The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
Cantero, Maria del Rocio
Cyclic Amp (Camp)
Electrophysiology
Placenta
Protein Kinase a (Pka)
title_short The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
title_full The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
title_fullStr The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
title_full_unstemmed The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
title_sort The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function
dc.creator.none.fl_str_mv Cantero, Maria del Rocio
Velázquez, Irina Florencia
Streets, Andrew J.
Ong, Albert C. M.
Cantiello, Horacio Fabio
author Cantero, Maria del Rocio
author_facet Cantero, Maria del Rocio
Velázquez, Irina Florencia
Streets, Andrew J.
Ong, Albert C. M.
Cantiello, Horacio Fabio
author_role author
author2 Velázquez, Irina Florencia
Streets, Andrew J.
Ong, Albert C. M.
Cantiello, Horacio Fabio
author2_role author
author
author
author
dc.subject.none.fl_str_mv Cyclic Amp (Camp)
Electrophysiology
Placenta
Protein Kinase a (Pka)
topic Cyclic Amp (Camp)
Electrophysiology
Placenta
Protein Kinase a (Pka)
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Polycystin-2 (PC2) is aTRP-type, Ca2+-permeable non-selective cation channel that plays an important role in Ca2+ signaling in renal and non-renal cells. The effect(s) of the cAMP pathway and kinase mediated phosphorylation of PC2 seem to be relevant to PC2 trafficking and its interaction with polycystin-1. However, the role of PC2 phosphorylation in channel function is still poorly defined. Here we reconstituted apical membranes of term human syncytiotrophoblast (hST), containing endogenous PC2 (PC2hst), and in vitro translated channel protein (PC2iv). Addition of the catalytic subunit of PKA increased by 566% the spontaneous PC2hst channel activity in the presence of ATP. Interestingly, 8-Br-cAMP also stimulated spontaneous PC2hst channel activity in the absence of the exogenous kinase. Either stimulation was inhibited by addition of alkaline phosphatase, which in turn, was reversed by the phosphatase inhibitor vanadate. Neither maneuver modified the single channel conductance but instead increased channel mean open time. PKA directly phosphorylated PC2, which increased the mean open time but not the single channel conductance of the channel. PKA phosphorylation did not modify either R742X truncated or S829A-mutant PC2iv channel function. The data indicate that the cAMP pathway regulates PC2-mediated cation transport in the hST. The relevant PKA site for PC2 channel regulation centers on a single residue serine 829, in the carboxyl terminus.
Fil: Cantero, Maria del Rocio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Velázquez, Irina Florencia. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Streets, Andrew J.. University of Sheffield Medical School; Reino Unido
Fil: Ong, Albert C. M.. University of Sheffield Medical School; Reino Unido
Fil: Cantiello, Horacio Fabio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Biofísica; Argentina
description Polycystin-2 (PC2) is aTRP-type, Ca2+-permeable non-selective cation channel that plays an important role in Ca2+ signaling in renal and non-renal cells. The effect(s) of the cAMP pathway and kinase mediated phosphorylation of PC2 seem to be relevant to PC2 trafficking and its interaction with polycystin-1. However, the role of PC2 phosphorylation in channel function is still poorly defined. Here we reconstituted apical membranes of term human syncytiotrophoblast (hST), containing endogenous PC2 (PC2hst), and in vitro translated channel protein (PC2iv). Addition of the catalytic subunit of PKA increased by 566% the spontaneous PC2hst channel activity in the presence of ATP. Interestingly, 8-Br-cAMP also stimulated spontaneous PC2hst channel activity in the absence of the exogenous kinase. Either stimulation was inhibited by addition of alkaline phosphatase, which in turn, was reversed by the phosphatase inhibitor vanadate. Neither maneuver modified the single channel conductance but instead increased channel mean open time. PKA directly phosphorylated PC2, which increased the mean open time but not the single channel conductance of the channel. PKA phosphorylation did not modify either R742X truncated or S829A-mutant PC2iv channel function. The data indicate that the cAMP pathway regulates PC2-mediated cation transport in the hST. The relevant PKA site for PC2 channel regulation centers on a single residue serine 829, in the carboxyl terminus.
publishDate 2015
dc.date.none.fl_str_mv 2015-09
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/11336/38777
Cantero, Maria del Rocio; Velázquez, Irina Florencia; Streets, Andrew J.; Ong, Albert C. M.; Cantiello, Horacio Fabio; The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 290; 39; 9-2015; 23888-23896
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/38777
identifier_str_mv Cantero, Maria del Rocio; Velázquez, Irina Florencia; Streets, Andrew J.; Ong, Albert C. M.; Cantiello, Horacio Fabio; The cAMP Signaling pathway and direct protein kinase a phosphorylation regulate polycystin-2 (TRPP2) Channel Function; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 290; 39; 9-2015; 23888-23896
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M115.661082
info:eu-repo/semantics/altIdentifier/url/http://www.jbc.org/content/290/39/23888.long
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4583051/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
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 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
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score 13.13397

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