title: AMP-activated protein kinase mediates VEGF-stimulated endothelial NO production.
abstract: Vascular endothelial growth factor (VEGF) is an important regulator of endothelial cell function. VEGF stimulates NO production, proposed to be a result of phosphorylation and activation of endothelial NO synthase (eNOS) at Ser1177. Phosphorylation of eNOS at this site also occurs after activation of AMP-activated protein kinase (AMPK) in cultured endothelial cells. We therefore determined whether AMPK mediates VEGF-stimulated NO synthesis in endothelial cells. VEGF caused a rapid, dose-dependent stimulation of AMPK activity, with a concomitant increase in phosphorylation of eNOS at Ser1177. Infection of endothelial cells with an adenovirus expressing a dominant negative mutant AMPK partially inhibited both VEGF-stimulated eNOS Ser1177 phosphorylation and NO production. VEGF-stimulated AMPK activity was completely inhibited by the Ca(2+)/calmodulin-dependent protein kinase kinase inhibitor, STO-609. Stimulation of AMPK via Ca(2+)/calmodulin-dependent protein kinase kinase represents a novel signalling mechanism utilised by VEGF in endothelial cells that contributes to eNOS phosphorylation and NO production.
authors.affiliation: Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, UK.
authors.forename: James A
authors.initials: JA
authors.lastname: Reihill
authors.forename: Marie-Ann
authors.initials: MA
authors.lastname: Ewart
authors.forename: D Grahame
authors.initials: DG
authors.lastname: Hardie
authors.forename: Ian P
authors.initials: IP
authors.lastname: Salt
mesh_terms: D055372:AMP-Activated Protein Kinases; D000595:Amino Acid Sequence; D000730:Androstadienes / Q000494:pharmacology; D001562:Benzimidazoles / Q000494:pharmacology; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D004730:Endothelium, Vascular / Q000502:physiology*; D004963:Estrenes / Q000494:pharmacology; D006801:Humans; D007546:Isoquinolines / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000502:physiology*; D053644:Naphthalimides / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D011760:Pyrrolidinones / Q000494:pharmacology; D012694:Serine / Q000378:metabolism; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D001562:Benzimidazoles; D004963:Estrenes; D007546:Isoquinolines; D009097:Multienzyme Complexes; D053644:Naphthalimides; D011760:Pyrrolidinones; C458525:STO 609; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D009569:Nitric Oxide; D012694:Serine; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D017346:Protein Serine-Threonine Kinases; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D055372:AMP-Activated Protein Kinases; D010738:Type C Phospholipases; D000077191:Wortmannin