Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G

Autores
Gonano, Luis Alberto; Aitken Buck, Hamish M.; Chakraborty, Akash D.; Worthington, Luke P. I.; Cully, Tanya R.; Lamberts, Regis R.; Vila Petroff, Martin Gerarde; Jones, Peter P.
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: The cGMP-dependent protein kinase G (PKG) phosphorylates the cardiac ryanodine receptor (RyR2) in vitro. We aimed to determine whether modulation of endogenous PKG alters RyR2-mediated spontaneous Ca2+ release and whether this effect is linked to a change in RyR2 phosphorylation. Methods: & Results: Human embryonic kidney (HEK293) cells with inducible RyR2 expression were treated with the cGMP analogue 8-Br-cGMP (100 μM) to activate endogenous PKG. In cells transfected with luminal Ca2+ sensor, D1ER, PKG activation significantly reduced the threshold for RyR2-mediated spontaneous Ca2+ release (93.9 ± 0.4% of store size with vehicle vs. 91.7 ± 0.8% with 8-Br-cGMP, P = 0.04). Mutation of the proposed PKG phosphorylation sites, S2808 and S2030, either individually or as a combination, prevented the decrease in Ca2+ release threshold induced by endogenous PKG activation. Interestingly, despite a functional dependence on expression of RyR2 phosphorylation sites, 8-Br-cGMP activation of PKG did not promote a detectable change in S2808 phosphorylation (P = 0.9). Paradoxically, pharmacological inhibition of PKG with KT 5823 (1 μM) also reduced the threshold for spontaneous Ca2+ release through RyR2 without affecting S2808 phosphorylation. Silencing RNA knockdown of endogenous PKG expression also had no quantifiable effect on RyR2 S2808 phosphorylation (P = 0.9). However, unlike PKG inhibition with KT 5823, PKG knockdown did not alter spontaneous Ca2+ release propensity or luminal Ca2+ handling. Conclusion: In an intact cell model, activation of endogenous PKG reduces the threshold for RyR2-mediated spontaneous Ca2+ release in a manner dependent on the RyR2 phosphorylation sites S2808 and S2030. This study clarifies the regulation of RyR2 Ca2+ release by endogenous PKG and functionally implicates the role of RyR2 phosphorylation.
Fil: Gonano, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentina. University of Otago; Nueva Zelanda
Fil: Aitken Buck, Hamish M.. University of Otago; Nueva Zelanda
Fil: Chakraborty, Akash D.. University of Otago; Nueva Zelanda
Fil: Worthington, Luke P. I.. University of Otago; Nueva Zelanda
Fil: Cully, Tanya R.. University of Otago; Nueva Zelanda
Fil: Lamberts, Regis R.. University of Otago; Nueva Zelanda
Fil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentina
Fil: Jones, Peter P.. University of Otago; Nueva Zelanda
Materia
CALCIUM
CARDIAC RYANODINE RECEPTOR
KT 5823
PHOSPHORYLATION
PROTEIN KINASE G
STORE OVERLOAD-INDUCED CALCIUM RELEASE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/209958
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/209958
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase GGonano, Luis AlbertoAitken Buck, Hamish M.Chakraborty, Akash D.Worthington, Luke P. I.Cully, Tanya R.Lamberts, Regis R.Vila Petroff, Martin GerardeJones, Peter P.CALCIUMCARDIAC RYANODINE RECEPTORKT 5823PHOSPHORYLATIONPROTEIN KINASE GSTORE OVERLOAD-INDUCED CALCIUM RELEASEhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Background: The cGMP-dependent protein kinase G (PKG) phosphorylates the cardiac ryanodine receptor (RyR2) in vitro. We aimed to determine whether modulation of endogenous PKG alters RyR2-mediated spontaneous Ca2+ release and whether this effect is linked to a change in RyR2 phosphorylation. Methods: & Results: Human embryonic kidney (HEK293) cells with inducible RyR2 expression were treated with the cGMP analogue 8-Br-cGMP (100 μM) to activate endogenous PKG. In cells transfected with luminal Ca2+ sensor, D1ER, PKG activation significantly reduced the threshold for RyR2-mediated spontaneous Ca2+ release (93.9 ± 0.4% of store size with vehicle vs. 91.7 ± 0.8% with 8-Br-cGMP, P = 0.04). Mutation of the proposed PKG phosphorylation sites, S2808 and S2030, either individually or as a combination, prevented the decrease in Ca2+ release threshold induced by endogenous PKG activation. Interestingly, despite a functional dependence on expression of RyR2 phosphorylation sites, 8-Br-cGMP activation of PKG did not promote a detectable change in S2808 phosphorylation (P = 0.9). Paradoxically, pharmacological inhibition of PKG with KT 5823 (1 μM) also reduced the threshold for spontaneous Ca2+ release through RyR2 without affecting S2808 phosphorylation. Silencing RNA knockdown of endogenous PKG expression also had no quantifiable effect on RyR2 S2808 phosphorylation (P = 0.9). However, unlike PKG inhibition with KT 5823, PKG knockdown did not alter spontaneous Ca2+ release propensity or luminal Ca2+ handling. Conclusion: In an intact cell model, activation of endogenous PKG reduces the threshold for RyR2-mediated spontaneous Ca2+ release in a manner dependent on the RyR2 phosphorylation sites S2808 and S2030. This study clarifies the regulation of RyR2 Ca2+ release by endogenous PKG and functionally implicates the role of RyR2 phosphorylation.Fil: Gonano, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentina. University of Otago; Nueva ZelandaFil: Aitken Buck, Hamish M.. University of Otago; Nueva ZelandaFil: Chakraborty, Akash D.. University of Otago; Nueva ZelandaFil: Worthington, Luke P. I.. University of Otago; Nueva ZelandaFil: Cully, Tanya R.. University of Otago; Nueva ZelandaFil: Lamberts, Regis R.. University of Otago; Nueva ZelandaFil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; ArgentinaFil: Jones, Peter P.. University of Otago; Nueva ZelandaElsevier2022-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/209958Gonano, Luis Alberto; Aitken Buck, Hamish M.; Chakraborty, Akash D.; Worthington, Luke P. I.; Cully, Tanya R.; et al.; Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G; Elsevier; Current Research in Physiology; 5; 1-2022; 171-1782665-9441CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S2665944122000165info:eu-repo/semantics/altIdentifier/doi/10.1016/j.crphys.2022.03.004info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:05:42Zoai:ri.conicet.gov.ar:11336/209958instacron: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:34982025-10-22 11:05:42.588CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
title Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
spellingShingle Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
Gonano, Luis Alberto
CALCIUM
CARDIAC RYANODINE RECEPTOR
KT 5823
PHOSPHORYLATION
PROTEIN KINASE G
STORE OVERLOAD-INDUCED CALCIUM RELEASE
title_short Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
title_full Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
title_fullStr Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
title_full_unstemmed Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
title_sort Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G
dc.creator.none.fl_str_mv Gonano, Luis Alberto
Aitken Buck, Hamish M.
Chakraborty, Akash D.
Worthington, Luke P. I.
Cully, Tanya R.
Lamberts, Regis R.
Vila Petroff, Martin Gerarde
Jones, Peter P.
author Gonano, Luis Alberto
author_facet Gonano, Luis Alberto
Aitken Buck, Hamish M.
Chakraborty, Akash D.
Worthington, Luke P. I.
Cully, Tanya R.
Lamberts, Regis R.
Vila Petroff, Martin Gerarde
Jones, Peter P.
author_role author
author2 Aitken Buck, Hamish M.
Chakraborty, Akash D.
Worthington, Luke P. I.
Cully, Tanya R.
Lamberts, Regis R.
Vila Petroff, Martin Gerarde
Jones, Peter P.
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CALCIUM
CARDIAC RYANODINE RECEPTOR
KT 5823
PHOSPHORYLATION
PROTEIN KINASE G
STORE OVERLOAD-INDUCED CALCIUM RELEASE
topic CALCIUM
CARDIAC RYANODINE RECEPTOR
KT 5823
PHOSPHORYLATION
PROTEIN KINASE G
STORE OVERLOAD-INDUCED CALCIUM RELEASE
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Background: The cGMP-dependent protein kinase G (PKG) phosphorylates the cardiac ryanodine receptor (RyR2) in vitro. We aimed to determine whether modulation of endogenous PKG alters RyR2-mediated spontaneous Ca2+ release and whether this effect is linked to a change in RyR2 phosphorylation. Methods: & Results: Human embryonic kidney (HEK293) cells with inducible RyR2 expression were treated with the cGMP analogue 8-Br-cGMP (100 μM) to activate endogenous PKG. In cells transfected with luminal Ca2+ sensor, D1ER, PKG activation significantly reduced the threshold for RyR2-mediated spontaneous Ca2+ release (93.9 ± 0.4% of store size with vehicle vs. 91.7 ± 0.8% with 8-Br-cGMP, P = 0.04). Mutation of the proposed PKG phosphorylation sites, S2808 and S2030, either individually or as a combination, prevented the decrease in Ca2+ release threshold induced by endogenous PKG activation. Interestingly, despite a functional dependence on expression of RyR2 phosphorylation sites, 8-Br-cGMP activation of PKG did not promote a detectable change in S2808 phosphorylation (P = 0.9). Paradoxically, pharmacological inhibition of PKG with KT 5823 (1 μM) also reduced the threshold for spontaneous Ca2+ release through RyR2 without affecting S2808 phosphorylation. Silencing RNA knockdown of endogenous PKG expression also had no quantifiable effect on RyR2 S2808 phosphorylation (P = 0.9). However, unlike PKG inhibition with KT 5823, PKG knockdown did not alter spontaneous Ca2+ release propensity or luminal Ca2+ handling. Conclusion: In an intact cell model, activation of endogenous PKG reduces the threshold for RyR2-mediated spontaneous Ca2+ release in a manner dependent on the RyR2 phosphorylation sites S2808 and S2030. This study clarifies the regulation of RyR2 Ca2+ release by endogenous PKG and functionally implicates the role of RyR2 phosphorylation.
Fil: Gonano, Luis Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentina. University of Otago; Nueva Zelanda
Fil: Aitken Buck, Hamish M.. University of Otago; Nueva Zelanda
Fil: Chakraborty, Akash D.. University of Otago; Nueva Zelanda
Fil: Worthington, Luke P. I.. University of Otago; Nueva Zelanda
Fil: Cully, Tanya R.. University of Otago; Nueva Zelanda
Fil: Lamberts, Regis R.. University of Otago; Nueva Zelanda
Fil: Vila Petroff, Martin Gerarde. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani". Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Investigaciones Cardiovasculares "Dr. Horacio Eugenio Cingolani"; Argentina
Fil: Jones, Peter P.. University of Otago; Nueva Zelanda
description Background: The cGMP-dependent protein kinase G (PKG) phosphorylates the cardiac ryanodine receptor (RyR2) in vitro. We aimed to determine whether modulation of endogenous PKG alters RyR2-mediated spontaneous Ca2+ release and whether this effect is linked to a change in RyR2 phosphorylation. Methods: & Results: Human embryonic kidney (HEK293) cells with inducible RyR2 expression were treated with the cGMP analogue 8-Br-cGMP (100 μM) to activate endogenous PKG. In cells transfected with luminal Ca2+ sensor, D1ER, PKG activation significantly reduced the threshold for RyR2-mediated spontaneous Ca2+ release (93.9 ± 0.4% of store size with vehicle vs. 91.7 ± 0.8% with 8-Br-cGMP, P = 0.04). Mutation of the proposed PKG phosphorylation sites, S2808 and S2030, either individually or as a combination, prevented the decrease in Ca2+ release threshold induced by endogenous PKG activation. Interestingly, despite a functional dependence on expression of RyR2 phosphorylation sites, 8-Br-cGMP activation of PKG did not promote a detectable change in S2808 phosphorylation (P = 0.9). Paradoxically, pharmacological inhibition of PKG with KT 5823 (1 μM) also reduced the threshold for spontaneous Ca2+ release through RyR2 without affecting S2808 phosphorylation. Silencing RNA knockdown of endogenous PKG expression also had no quantifiable effect on RyR2 S2808 phosphorylation (P = 0.9). However, unlike PKG inhibition with KT 5823, PKG knockdown did not alter spontaneous Ca2+ release propensity or luminal Ca2+ handling. Conclusion: In an intact cell model, activation of endogenous PKG reduces the threshold for RyR2-mediated spontaneous Ca2+ release in a manner dependent on the RyR2 phosphorylation sites S2808 and S2030. This study clarifies the regulation of RyR2 Ca2+ release by endogenous PKG and functionally implicates the role of RyR2 phosphorylation.
publishDate 2022
dc.date.none.fl_str_mv 2022-01
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/11336/209958
Gonano, Luis Alberto; Aitken Buck, Hamish M.; Chakraborty, Akash D.; Worthington, Luke P. I.; Cully, Tanya R.; et al.; Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G; Elsevier; Current Research in Physiology; 5; 1-2022; 171-178
2665-9441
CONICET Digital
CONICET
url http://hdl.handle.net/11336/209958
identifier_str_mv Gonano, Luis Alberto; Aitken Buck, Hamish M.; Chakraborty, Akash D.; Worthington, Luke P. I.; Cully, Tanya R.; et al.; Regulation of cardiac ryanodine receptor function by the cyclic-GMP dependent protein kinase G; Elsevier; Current Research in Physiology; 5; 1-2022; 171-178
2665-9441
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://linkinghub.elsevier.com/retrieve/pii/S2665944122000165
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.crphys.2022.03.004
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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 12.982451

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