PMID: 9194575May 27, 1997Paper

The thrombin receptor in adrenal medullary microvascular endothelial cells is negatively coupled to adenylyl cyclase through a Gi protein

Biochimica Et Biophysica Acta
V G ManolopoulosP I Lelkes

Abstract

The effects of thrombin on adenylyl cyclase activity were examined in rat adrenal medullary microvascular endothelial cells (RAMEC). Confluent RAMEC monolayers were stimulated for 5 min with cAMP-generating agents in the absence and presence of thrombin, and intracellular cAMP was measured with a radioligand binding assay. Thrombin (0.001-0.25 U/ml) dose-dependently inhibited IBMX-, isoproterenol- and forskolin-stimulated cAMP accumulation. A peptide agonist of the thrombin receptor, gamma-thrombin, and the serine proteases trypsin and plasmin, also inhibited agonist-stimulated cAMP levels, while proteolytically inactive PPACK- or DIP-alpha-thrombins were without effect. Moreover, the thrombin inhibitor hirudin abolished the inhibitory effect of thrombin but not of the peptide agonist. These results suggest that the inhibitory action of thrombin on cAMP accumulation is mediated by a proteolytically-activated thrombin receptor. The inhibitor of G(i)-proteins pertussis toxin abolished the inhibitory effect of thrombin on isoproterenol- or IBMX-stimulated cAMP production, while the phorbol ester PMA partly impaired it. The protein kinase C inhibitors staurosporine or H7 and the intracellular Ca2+ chelator BAPTA-AM were without eff...Continue Reading

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Citations

Jan 8, 2004·Microvascular Research·Anna A BirukovaAlexander D Verin
Sep 7, 2000·General Pharmacology·V G ManolopoulosB Nilius
Mar 6, 2002·British Journal of Pharmacology·Samuel AsfahaNathalie Vergnolle
Mar 21, 2002·Science's STKE : Signal Transduction Knowledge Environment·Darren G Woodside
Dec 4, 2004·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Anna A BirukovaAlexander D Verin
Dec 24, 2005·Physiological Reviews·Dolly Mehta, Asrar B Malik
Dec 10, 2002·American Journal of Physiology. Lung Cellular and Molecular Physiology·Judy R CreightonTroy Stevens
Feb 28, 2006·British Journal of Pharmacology·Maurizio BifulcoPatrizia Gazzerro
Feb 24, 2001·Endothelium : Journal of Endothelial Cell Research·C E PattersonJ G Garcia
Jun 21, 2001·Endothelium : Journal of Endothelial Cell Research· Burnatowska-HledinC Barney
Sep 27, 2001·Endothelium : Journal of Endothelial Cell Research·C E Patterson, H Lum
Feb 5, 2002·Endothelium : Journal of Endothelial Cell Research·E PapadimitriouP L Lelkes
Jul 3, 1999·Physiological Reviews·C C Michel, F E Curry
Feb 5, 2002·Endothelium : Journal of Endothelial Cell Research·E PapadimitriouP I Lelkes
Jun 7, 2011·Microvascular Research·Tracy Thennes, Dolly Mehta

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