Protein kinase A-dependent phosphorylation of ryanodine receptors increases Ca2+ leak in mouse heart

Satoshi Morimoto; Jin O-Uchi; Makoto Kawai; Toshiyuki Hoshina; Yoichiro Kusakari; Kimiaki Komukai; Hiroyuki Sasaki; Kenichi Hongo; Satoshi Kurihara

doi:10.1016/j.bbrc.2009.09.071

Protein kinase A-dependent phosphorylation of ryanodine receptors increases Ca²⁺ leak in mouse heart

Satoshi Morimoto, Jin O-Uchi, Makoto Kawai, Toshiyuki Hoshina, Yoichiro Kusakari, Kimiaki Komukai, Hiroyuki Sasaki, Kenichi Hongo, Satoshi Kurihara

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

In heart failure, chronic catecholaminergic stimulation increases diastolic Ca²⁺ leak from ryanodine receptors (RyRs) of sarcoplasmic reticulum (SR), possibly due to the phosphorylation of RyRs through the activation of protein kinase A (PKA) or Ca²⁺/calmodulin-dependent protein kinase II (CaMKII). In the present study, we attempted to identify which activated kinase is responsible for the enhanced Ca²⁺ leak caused by β-adrenergic stimulation. Trabeculae obtained from the hearts of adult male C57BL/6J mice were treated with isoproterenol and then permeabilized with saponin. To examine SR functions, Ca²⁺ in SR was released with caffeine and measured with fluo-3. The Ca²⁺ leak in isoproterenol-treated preparations was significantly increased when the PKA-dependent phosphorylation of RyR was increased without the involvement of CaMKII-dependent phosphorylation. Both the increase in Ca²⁺ leak and the phosphorylation of RyR were blocked by a PKA inhibitor. Our results show that β-adrenergic stimulation increases Ca²⁺ leak from SR through PKA-dependent phosphorylation of RyR.

Original language	English (US)
Pages (from-to)	87-92
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	390
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2009.09.071
State	Published - Dec 4 2009
Externally published	Yes

Bibliographical note

Funding Information:
Source of Funding: This study was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology [ 18590789 to K.K., 19590833 to K.H., and 19500357 S.K.], the Uehara Memorial Foundation [to Y.K. and S.K.], the Takeda Science Foundation [to K.H.], the Vehicle Racing Commemorative Foundation [to K.H. and S.K.], a Japan Heart Foundation Young Investigator’s Research Grant [to J.O.-U.], the Japan Foundation of Cardiovascular Research [to J.O.-U.], the Kato Memorial Bioscience Foundation [to J.O.-U.], and The Jikei University Research Fund [to J.O.-U.].

Keywords

Ca leak
Ca/calmodulin-dependent protein kinase II
Protein kinase A
Ryanodine receptor
β-Adrenoceptor stimulation

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@article{3c75af60c0854c208f2b98013e6baaef,

title = "Protein kinase A-dependent phosphorylation of ryanodine receptors increases Ca2+ leak in mouse heart",

abstract = "In heart failure, chronic catecholaminergic stimulation increases diastolic Ca2+ leak from ryanodine receptors (RyRs) of sarcoplasmic reticulum (SR), possibly due to the phosphorylation of RyRs through the activation of protein kinase A (PKA) or Ca2+/calmodulin-dependent protein kinase II (CaMKII). In the present study, we attempted to identify which activated kinase is responsible for the enhanced Ca2+ leak caused by β-adrenergic stimulation. Trabeculae obtained from the hearts of adult male C57BL/6J mice were treated with isoproterenol and then permeabilized with saponin. To examine SR functions, Ca2+ in SR was released with caffeine and measured with fluo-3. The Ca2+ leak in isoproterenol-treated preparations was significantly increased when the PKA-dependent phosphorylation of RyR was increased without the involvement of CaMKII-dependent phosphorylation. Both the increase in Ca2+ leak and the phosphorylation of RyR were blocked by a PKA inhibitor. Our results show that β-adrenergic stimulation increases Ca2+ leak from SR through PKA-dependent phosphorylation of RyR.",

keywords = "Ca leak, Ca/calmodulin-dependent protein kinase II, Protein kinase A, Ryanodine receptor, β-Adrenoceptor stimulation",

author = "Satoshi Morimoto and Jin O-Uchi and Makoto Kawai and Toshiyuki Hoshina and Yoichiro Kusakari and Kimiaki Komukai and Hiroyuki Sasaki and Kenichi Hongo and Satoshi Kurihara",

note = "Funding Information: Source of Funding: This study was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology [ 18590789 to K.K., 19590833 to K.H., and 19500357 S.K.], the Uehara Memorial Foundation [to Y.K. and S.K.], the Takeda Science Foundation [to K.H.], the Vehicle Racing Commemorative Foundation [to K.H. and S.K.], a Japan Heart Foundation Young Investigator{\textquoteright}s Research Grant [to J.O.-U.], the Japan Foundation of Cardiovascular Research [to J.O.-U.], the Kato Memorial Bioscience Foundation [to J.O.-U.], and The Jikei University Research Fund [to J.O.-U.]. ",

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AU - Morimoto, Satoshi

AU - O-Uchi, Jin

AU - Kawai, Makoto

AU - Hoshina, Toshiyuki

AU - Kusakari, Yoichiro

AU - Komukai, Kimiaki

AU - Sasaki, Hiroyuki

AU - Hongo, Kenichi

AU - Kurihara, Satoshi

N1 - Funding Information: Source of Funding: This study was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology [ 18590789 to K.K., 19590833 to K.H., and 19500357 S.K.], the Uehara Memorial Foundation [to Y.K. and S.K.], the Takeda Science Foundation [to K.H.], the Vehicle Racing Commemorative Foundation [to K.H. and S.K.], a Japan Heart Foundation Young Investigator’s Research Grant [to J.O.-U.], the Japan Foundation of Cardiovascular Research [to J.O.-U.], the Kato Memorial Bioscience Foundation [to J.O.-U.], and The Jikei University Research Fund [to J.O.-U.].

PY - 2009/12/4

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N2 - In heart failure, chronic catecholaminergic stimulation increases diastolic Ca2+ leak from ryanodine receptors (RyRs) of sarcoplasmic reticulum (SR), possibly due to the phosphorylation of RyRs through the activation of protein kinase A (PKA) or Ca2+/calmodulin-dependent protein kinase II (CaMKII). In the present study, we attempted to identify which activated kinase is responsible for the enhanced Ca2+ leak caused by β-adrenergic stimulation. Trabeculae obtained from the hearts of adult male C57BL/6J mice were treated with isoproterenol and then permeabilized with saponin. To examine SR functions, Ca2+ in SR was released with caffeine and measured with fluo-3. The Ca2+ leak in isoproterenol-treated preparations was significantly increased when the PKA-dependent phosphorylation of RyR was increased without the involvement of CaMKII-dependent phosphorylation. Both the increase in Ca2+ leak and the phosphorylation of RyR were blocked by a PKA inhibitor. Our results show that β-adrenergic stimulation increases Ca2+ leak from SR through PKA-dependent phosphorylation of RyR.

AB - In heart failure, chronic catecholaminergic stimulation increases diastolic Ca2+ leak from ryanodine receptors (RyRs) of sarcoplasmic reticulum (SR), possibly due to the phosphorylation of RyRs through the activation of protein kinase A (PKA) or Ca2+/calmodulin-dependent protein kinase II (CaMKII). In the present study, we attempted to identify which activated kinase is responsible for the enhanced Ca2+ leak caused by β-adrenergic stimulation. Trabeculae obtained from the hearts of adult male C57BL/6J mice were treated with isoproterenol and then permeabilized with saponin. To examine SR functions, Ca2+ in SR was released with caffeine and measured with fluo-3. The Ca2+ leak in isoproterenol-treated preparations was significantly increased when the PKA-dependent phosphorylation of RyR was increased without the involvement of CaMKII-dependent phosphorylation. Both the increase in Ca2+ leak and the phosphorylation of RyR were blocked by a PKA inhibitor. Our results show that β-adrenergic stimulation increases Ca2+ leak from SR through PKA-dependent phosphorylation of RyR.

KW - Ca leak

KW - Ca/calmodulin-dependent protein kinase II

KW - Protein kinase A

KW - Ryanodine receptor

KW - β-Adrenoceptor stimulation

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