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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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AMP-Activated Protein Kinases
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Amino Acid Sequence
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Androstadienes / pharmacology
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Benzimidazoles / pharmacology
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Calcium-Calmodulin-Dependent Protein Kinase Kinase
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Endothelium, Vascular / physiology*
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Estrenes / pharmacology
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Humans
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Isoquinolines / pharmacology
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Multienzyme Complexes / physiology*
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Naphthalimides / pharmacology
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Nitric Oxide / biosynthesis*
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Nitric Oxide Synthase Type III / metabolism
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Protein Serine-Threonine Kinases / antagonists & inhibitors
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Protein Serine-Threonine Kinases / physiology*
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Pyrrolidinones / pharmacology
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Serine / metabolism
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Type C Phospholipases / antagonists & inhibitors
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Vascular Endothelial Growth Factor A / metabolism*
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Wortmannin
Substances
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Androstadienes
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Benzimidazoles
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Estrenes
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Isoquinolines
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Multienzyme Complexes
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Naphthalimides
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Pyrrolidinones
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STO 609
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VEGFA protein, human
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Vascular Endothelial Growth Factor A
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1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
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Nitric Oxide
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Serine
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NOS3 protein, human
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Nitric Oxide Synthase Type III
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Protein Serine-Threonine Kinases
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Calcium-Calmodulin-Dependent Protein Kinase Kinase
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AMP-Activated Protein Kinases
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Type C Phospholipases
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Wortmannin