Ca<SUP>2+</SUP>/calmodulin kinase II increases ryanodine binding and Ca<SUP>2+</SUP>-induced sarcoplasmic reticulum Ca<SUP>2+</SUP> release kinetics during beta-adrenergic stimulation

We aimed to define the relative contribution of both PKA and Ca2+/calmodulin-dependent protein kinase II (CaMKII) cascades to the phosphorylation of RyR2 and the activity of the channel during β-adrenergic receptor (βAR) stimulation. Rat hearts were perfused with increasing concentrations of the β-a...

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Autores principales: Ferrero, Paola Viviana, Said, María Matilde, Sánchez, Gina, Vittone, Leticia, Valverde, Carlos Alfredo, Donoso, Paulina, Mattiazzi, Alicia Ramona, Mundiña-Weilenmann, Cecilia
Formato: Articulo Preprint
Lenguaje:Inglés
Publicado: 2007
Materias:
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/108000
http://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC2045504&blobtype=pdf
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id I19-R120-10915-108000
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Médicas
Ryanodine receptor
β-adrenergic stimulation
CaMKII-dependent phosphorylation
Sarcoplasmic reticulum
Ca2+ release
spellingShingle Ciencias Médicas
Ryanodine receptor
β-adrenergic stimulation
CaMKII-dependent phosphorylation
Sarcoplasmic reticulum
Ca2+ release
Ferrero, Paola Viviana
Said, María Matilde
Sánchez, Gina
Vittone, Leticia
Valverde, Carlos Alfredo
Donoso, Paulina
Mattiazzi, Alicia Ramona
Mundiña-Weilenmann, Cecilia
Ca<SUP>2+</SUP>/calmodulin kinase II increases ryanodine binding and Ca<SUP>2+</SUP>-induced sarcoplasmic reticulum Ca<SUP>2+</SUP> release kinetics during beta-adrenergic stimulation
topic_facet Ciencias Médicas
Ryanodine receptor
β-adrenergic stimulation
CaMKII-dependent phosphorylation
Sarcoplasmic reticulum
Ca2+ release
description We aimed to define the relative contribution of both PKA and Ca2+/calmodulin-dependent protein kinase II (CaMKII) cascades to the phosphorylation of RyR2 and the activity of the channel during β-adrenergic receptor (βAR) stimulation. Rat hearts were perfused with increasing concentrations of the β-agonist isoproterenol in the absence and the presence of CaMKII inhibition. CaMKII was inhibited either by preventing the Ca2+ influx to the cell by low [Ca]o plus nifedipine or by the specific inhibitor KN-93. We immunodetected RyR2 phosphorylated at Ser2809 (PKA and putative CaMKII site) and at Ser2815 (CaMKII site) and measured [3H]-ryanodine binding and fast Ca2+ release kinetics in sarcoplasmic reticulum (SR) vesicles. SR vesicles were isolated in conditions that preserved the phosphorylation levels achieved in the intact heart and were actively and equally loaded with Ca2+. Our results demonstrated that Ser2809 and Ser2815 of RyR2 were dose-dependently phosphorylated under βAR stimulation by PKA and CaMKII, respectively. The isoproterenolinduced increase in the phosphorylation of Ser2815 site was prevented by the PKA inhibitor H-89 and mimicked by forskolin. CaMKII-dependent phosphorylation of RyR2 (but not PKA-dependent phosphorylation) was responsible for the β-induced increase in the channel activity as indicated by the enhancement of the [3H]-ryanodine binding and the velocity of fast SR Ca2+ release. The present results show for the first time a dose-dependent increase in the phosphorylation of Ser2815 of RyR2 through the PKA-dependent activation of CaMKII and a predominant role of CaMKII-dependent phosphorylation of RyR2, over that of PKA-dependent phosphorylation, on SR-Ca2+ release during βAR stimulation.
format Articulo
Preprint
author Ferrero, Paola Viviana
Said, María Matilde
Sánchez, Gina
Vittone, Leticia
Valverde, Carlos Alfredo
Donoso, Paulina
Mattiazzi, Alicia Ramona
Mundiña-Weilenmann, Cecilia
author_facet Ferrero, Paola Viviana
Said, María Matilde
Sánchez, Gina
Vittone, Leticia
Valverde, Carlos Alfredo
Donoso, Paulina
Mattiazzi, Alicia Ramona
Mundiña-Weilenmann, Cecilia
author_sort Ferrero, Paola Viviana
title Ca<SUP>2+</SUP>/calmodulin kinase II increases ryanodine binding and Ca<SUP>2+</SUP>-induced sarcoplasmic reticulum Ca<SUP>2+</SUP> release kinetics during beta-adrenergic stimulation
title_short Ca<SUP>2+</SUP>/calmodulin kinase II increases ryanodine binding and Ca<SUP>2+</SUP>-induced sarcoplasmic reticulum Ca<SUP>2+</SUP> release kinetics during beta-adrenergic stimulation
title_full Ca<SUP>2+</SUP>/calmodulin kinase II increases ryanodine binding and Ca<SUP>2+</SUP>-induced sarcoplasmic reticulum Ca<SUP>2+</SUP> release kinetics during beta-adrenergic stimulation
title_fullStr Ca<SUP>2+</SUP>/calmodulin kinase II increases ryanodine binding and Ca<SUP>2+</SUP>-induced sarcoplasmic reticulum Ca<SUP>2+</SUP> release kinetics during beta-adrenergic stimulation
title_full_unstemmed Ca<SUP>2+</SUP>/calmodulin kinase II increases ryanodine binding and Ca<SUP>2+</SUP>-induced sarcoplasmic reticulum Ca<SUP>2+</SUP> release kinetics during beta-adrenergic stimulation
title_sort ca<sup>2+</sup>/calmodulin kinase ii increases ryanodine binding and ca<sup>2+</sup>-induced sarcoplasmic reticulum ca<sup>2+</sup> release kinetics during beta-adrenergic stimulation
publishDate 2007
url http://sedici.unlp.edu.ar/handle/10915/108000
http://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC2045504&blobtype=pdf
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