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Modulation of protein kinase C by adenosine: Involvement of adenosine A1 receptor-pertussis toxin sensitive nucleotide binding protein system

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Abstract

The objective of this study was to determine whether adenosine A1 or A2 receptor was responsible for the regulation of protein kinase C (PKC) in porcine coronary artery and its coupling to G-protein. Endothelium denuded arterial rings were incubated with PDBu (200nM) in the presence or absence of adenosine receptor agonists and antagonists for 1 day. Following incubation, the arterial rings were contracted with increasing concentrations of endothelin-1 (ET-1) (10−10−10−7M). Arteries incubated with PDBu alone failed to produce contraction in response to ET-1. On the contrary, inclusion of A1 receptor agonist ENBA at 10−9M in the incubation media with PDBu protected against the PDBu induced blunting of the ET-1 contractions by 50%. Incubation with ENBA alone increased ET-1 dependent contractions by about 2 fold. Inclusion of A1 receptor antagonist, N0861 at 10−6 M along with PDBu and ENBA, completely blocked the protective effect of ENBA against the PDBu induced attenuation of ET-1 contractions. N0861 also completely blocked the increase in ET-1 contractions in the arterial rings incubated with ENBA alone. Another A1 receptor antagonist DPCPX also produced similar results as N0861. On the contrary, arterial rings incubated with relatively specific A2 receptor agonist CGS 21680 at 10−4M did not produce any protection against PDBu induced blunting of the ET-1 contractions. Incubation with CGS 21680 alone also did not significantly alter the ET-1 contractions. Interestingly, inclusion of A2 receptor antagonist DMPX at 10−4M in the incubation media along with CGS 21680 mimicked the effects of ENBA alone i.e. produced protection against PDBu and enhanced ET-1 contractions. Incubation of the arteries with ENBA alone caused an accumulation of PKC levels, whereas, incubation with CGS 21680 had no significant effect on PKC levels. To study the coupling of adenosine receptor with G-protein, the tissue was incubated for one day with cholera (CT) or pertussis toxin (PT) in the presence or absence or ENBA and PDBu as described above. Incubation with PT blocked the protective effect of ENBA against PDBu as well as the elevation of ET-1 response when incubated with ENBA alone. On the contrary, incubation with CT did not produce any significant effect on ENBA responses. These results indicate that PKC is modulated by adenosine via A1 adenosine receptors and through a PT sensitive G-protein.

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Abbreviations

A2-R:

adenosine A2 receptor

Gs :

stimulatory guanine nucleotide binding protein

AC:

adenylate cyclase

A1-R:

adenosine A1 receptor

Gi :

inhibitory guanine nucleotide binding protein

P:

protein kinase C specific protease

PKC:

protein kinase C

ET-1 R:

endothelin-1 receptor

G:

guanine nucleotide binding protein

PLC:

phospholipase C

PIP2 :

phosphatidylinositol 4,5-bisphosphate

DAG:

diacylglycerol

IP3 :

d-myo-inositol 1,4,5-triphosphate

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Authors and Affiliations

  1. Department of Pharmacology, School of Medicine, East Carolina University, 27858, Greenville, NC, USA

    Ravi B. Marala & S. Jamal Mustafa

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  1. Ravi B. Marala
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  2. S. Jamal Mustafa
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This work was supported by National Heart, Lung and Blood Institute Grant HL-27339.

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Marala, R.B., Mustafa, S.J. Modulation of protein kinase C by adenosine: Involvement of adenosine A1 receptor-pertussis toxin sensitive nucleotide binding protein system. Mol Cell Biochem 149, 51–58 (1995). https://doi.org/10.1007/BF01076563

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