Adenosine A1 receptors selectively target protein kinase C isoforms to the caveolin-rich plasma membrane in cardiac myocytes

Zhaogang Yang, Wei Sun, Keli Hu

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Adenosine is a naturally occurring nucleoside that has been shown to regulate a variety of functions in the cardiovascular system. However, the mechanisms in adenosine receptor signaling are not completely understood. Given that adenosine receptors have been linked to protein kinase C (PKC) in cardioprotection and caveolae is critical for receptor signaling, we sought to determine whether activation of adenosine A1 receptors induces selective translocation of PKC isoforms to the membrane from the cytosol and whether activated PKC is targeted to the caveolin-rich plasma membrane microdomains. The freshly isolated adult rat cardiac myocytes were used to examine PKC isoforms including PKCα, PKCβ, PKCe{open}, PKCδ and PKCζ. Immunoblot analysis revealed that the immunoreactivity for PKCe{open} or PKCδ but not for PKCα, PKCβ or PKCζ increased significantly in the membrane fractions from cells pretreated with the selective adenosine A1 receptor agonist 2-chloro-N(6)-cyclopentyladenosine (CCPA, 100 nM) when compared with non-stimulated cells. The effect of CCPA on PKCe{open} or PKCδ translocation was blocked by adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 100 nM). When Western blot was performed from the caveolin-enriched plasma membrane fractions, the immunoreactivity for PKCe{open} or PKCδ but not PKCα, PKCβ or PKCζ was enhanced significantly by CCPA. Furthermore, PKCe{open} and PKCδ were detected in the anti-caveolin-3 immunoprecipitates but not in the samples without primary antibody. Immunofluorescence staining further indicates increased colocalization of PKCe{open} or PKCδ with caveolin-3 at cell peripheral region and T-tubular-like structures in response to adenosine A1 receptor activation. In conclusion, we demonstrate that activation of adenosine A1 receptors promotes the selective translocation of PKCe{open} and PKCδ to the caveolin-enriched plasma membrane microdomains in cardiac myocytes.

Original languageEnglish (US)
Pages (from-to)1868-1875
Number of pages8
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1793
Issue number12
DOIs
StatePublished - Dec 1 2009
Externally publishedYes

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Caveolins
Adenosine A1 Receptors
Cardiac Myocytes
Protein Kinase C
Protein Isoforms
Cell Membrane
Caveolin 3
Membrane Microdomains
2-chloro-N(6)cyclopentyladenosine
Purinergic P1 Receptors
Adenosine A1 Receptor Agonists
Adenosine A1 Receptor Antagonists
Caveolae

Keywords

  • Adenosine receptor
  • Cardiac myocyte
  • Caveolae
  • Caveolin-3
  • Protein kinase c

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

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title = "Adenosine A1 receptors selectively target protein kinase C isoforms to the caveolin-rich plasma membrane in cardiac myocytes",
abstract = "Adenosine is a naturally occurring nucleoside that has been shown to regulate a variety of functions in the cardiovascular system. However, the mechanisms in adenosine receptor signaling are not completely understood. Given that adenosine receptors have been linked to protein kinase C (PKC) in cardioprotection and caveolae is critical for receptor signaling, we sought to determine whether activation of adenosine A1 receptors induces selective translocation of PKC isoforms to the membrane from the cytosol and whether activated PKC is targeted to the caveolin-rich plasma membrane microdomains. The freshly isolated adult rat cardiac myocytes were used to examine PKC isoforms including PKCα, PKCβ, PKCe{open}, PKCδ and PKCζ. Immunoblot analysis revealed that the immunoreactivity for PKCe{open} or PKCδ but not for PKCα, PKCβ or PKCζ increased significantly in the membrane fractions from cells pretreated with the selective adenosine A1 receptor agonist 2-chloro-N(6)-cyclopentyladenosine (CCPA, 100 nM) when compared with non-stimulated cells. The effect of CCPA on PKCe{open} or PKCδ translocation was blocked by adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 100 nM). When Western blot was performed from the caveolin-enriched plasma membrane fractions, the immunoreactivity for PKCe{open} or PKCδ but not PKCα, PKCβ or PKCζ was enhanced significantly by CCPA. Furthermore, PKCe{open} and PKCδ were detected in the anti-caveolin-3 immunoprecipitates but not in the samples without primary antibody. Immunofluorescence staining further indicates increased colocalization of PKCe{open} or PKCδ with caveolin-3 at cell peripheral region and T-tubular-like structures in response to adenosine A1 receptor activation. In conclusion, we demonstrate that activation of adenosine A1 receptors promotes the selective translocation of PKCe{open} and PKCδ to the caveolin-enriched plasma membrane microdomains in cardiac myocytes.",
keywords = "Adenosine receptor, Cardiac myocyte, Caveolae, Caveolin-3, Protein kinase c",
author = "Zhaogang Yang and Wei Sun and Keli Hu",
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T1 - Adenosine A1 receptors selectively target protein kinase C isoforms to the caveolin-rich plasma membrane in cardiac myocytes

AU - Yang, Zhaogang

AU - Sun, Wei

AU - Hu, Keli

PY - 2009/12/1

Y1 - 2009/12/1

N2 - Adenosine is a naturally occurring nucleoside that has been shown to regulate a variety of functions in the cardiovascular system. However, the mechanisms in adenosine receptor signaling are not completely understood. Given that adenosine receptors have been linked to protein kinase C (PKC) in cardioprotection and caveolae is critical for receptor signaling, we sought to determine whether activation of adenosine A1 receptors induces selective translocation of PKC isoforms to the membrane from the cytosol and whether activated PKC is targeted to the caveolin-rich plasma membrane microdomains. The freshly isolated adult rat cardiac myocytes were used to examine PKC isoforms including PKCα, PKCβ, PKCe{open}, PKCδ and PKCζ. Immunoblot analysis revealed that the immunoreactivity for PKCe{open} or PKCδ but not for PKCα, PKCβ or PKCζ increased significantly in the membrane fractions from cells pretreated with the selective adenosine A1 receptor agonist 2-chloro-N(6)-cyclopentyladenosine (CCPA, 100 nM) when compared with non-stimulated cells. The effect of CCPA on PKCe{open} or PKCδ translocation was blocked by adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 100 nM). When Western blot was performed from the caveolin-enriched plasma membrane fractions, the immunoreactivity for PKCe{open} or PKCδ but not PKCα, PKCβ or PKCζ was enhanced significantly by CCPA. Furthermore, PKCe{open} and PKCδ were detected in the anti-caveolin-3 immunoprecipitates but not in the samples without primary antibody. Immunofluorescence staining further indicates increased colocalization of PKCe{open} or PKCδ with caveolin-3 at cell peripheral region and T-tubular-like structures in response to adenosine A1 receptor activation. In conclusion, we demonstrate that activation of adenosine A1 receptors promotes the selective translocation of PKCe{open} and PKCδ to the caveolin-enriched plasma membrane microdomains in cardiac myocytes.

AB - Adenosine is a naturally occurring nucleoside that has been shown to regulate a variety of functions in the cardiovascular system. However, the mechanisms in adenosine receptor signaling are not completely understood. Given that adenosine receptors have been linked to protein kinase C (PKC) in cardioprotection and caveolae is critical for receptor signaling, we sought to determine whether activation of adenosine A1 receptors induces selective translocation of PKC isoforms to the membrane from the cytosol and whether activated PKC is targeted to the caveolin-rich plasma membrane microdomains. The freshly isolated adult rat cardiac myocytes were used to examine PKC isoforms including PKCα, PKCβ, PKCe{open}, PKCδ and PKCζ. Immunoblot analysis revealed that the immunoreactivity for PKCe{open} or PKCδ but not for PKCα, PKCβ or PKCζ increased significantly in the membrane fractions from cells pretreated with the selective adenosine A1 receptor agonist 2-chloro-N(6)-cyclopentyladenosine (CCPA, 100 nM) when compared with non-stimulated cells. The effect of CCPA on PKCe{open} or PKCδ translocation was blocked by adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 100 nM). When Western blot was performed from the caveolin-enriched plasma membrane fractions, the immunoreactivity for PKCe{open} or PKCδ but not PKCα, PKCβ or PKCζ was enhanced significantly by CCPA. Furthermore, PKCe{open} and PKCδ were detected in the anti-caveolin-3 immunoprecipitates but not in the samples without primary antibody. Immunofluorescence staining further indicates increased colocalization of PKCe{open} or PKCδ with caveolin-3 at cell peripheral region and T-tubular-like structures in response to adenosine A1 receptor activation. In conclusion, we demonstrate that activation of adenosine A1 receptors promotes the selective translocation of PKCe{open} and PKCδ to the caveolin-enriched plasma membrane microdomains in cardiac myocytes.

KW - Adenosine receptor

KW - Cardiac myocyte

KW - Caveolae

KW - Caveolin-3

KW - Protein kinase c

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