P2 purinoceptor-stimulated conversion of arginine to citrulline in bovine endothelial cells is reduced by inhibition of protein kinase C

Biochemical Pharmacology
C BrownM R Boarder

Abstract

Bovine aortic endothelial cells contain two coexisting receptors for extracellular ATP, named the P2Y and P2U purinoceptors. Previous studies have shown that these receptors are linked to phospholipase C in a manner that is modulated in part by protein kinase C (PKC). In this study, we investigate the influence of PKC in the regulation of endothelial nitric oxide synthase (NOS) by these two purinoceptors. Activation of either P2Y or P2U purinoceptors by either 2-methylthio-ATP or UTP, respectively, stimulated the formation of [3H]-citrulline in [3H]-arginine-labelled cells in a concentration-dependent manner. This stimulation was sensitive to inhibition by NG-nitro-L-arginine. Ten minutes of pretreatment with the PKC activator tetradecanoyl phorbol acetate (TPA) failed to affect NOS activity, either alone or when stimulated with 2-methylthio-ATP or UTP. However, under these conditions TPA caused almost complete translocation of PKC-alpha from the cytosol to the membrane. Ten minutes of pretreatment with the PKC inhibitor Ro 31-8220 significantly inhibited the agonist-induced stimulation of NOS. These results show that both P2Y and P2U purinoceptors stimulate endothelial NOS in a manner that is dependent on PKC activity.

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Citations

May 19, 1998·Trends in Pharmacological Sciences·M R Boarder, S M Hourani
Mar 30, 2010·Acta Physiologica·G BurnstockA Verkhratsky
Jan 15, 2008·American Journal of Physiology. Lung Cellular and Molecular Physiology·Neetu SudStephen M Black
Feb 14, 2004·American Journal of Physiology. Heart and Circulatory Physiology·Girija G KonduriAshwani K Khanna
Aug 18, 2006·Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology·Mercedes MontielEugenio Jiménez
Aug 5, 1998·The American Journal of Physiology·A F CastroA C Taquini
Sep 18, 2002·American Journal of Physiology. Heart and Circulatory Physiology·Girija G Konduri, Janine Mattei
Mar 29, 2003·American Journal of Physiology. Heart and Circulatory Physiology·Girija G KonduriKirkwood A Pritchard
Oct 7, 2004·European Journal of Pharmacology·Zdenĕk ZídekAntonín Holý

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