Regulation of endothelium-derived nitric oxide production by the protein kinase Akt.
Endothelial nitric oxide synthase (eNOS) is the nitric oxide synthase isoform responsible for maintaining systemic blood pressure, vascular remodelling and angiogenesis. eNOS is phosphorylated in response to various forms of cellular stimulation, but the role of phosphorylation in the regulation of nitric oxide (NO) production and the kinase(s) responsible are not known. Here we show that the serine/threonine protein kinase Akt (protein kinase B) can directly phosphorylate eNOS on serine 1179 and activate the enzyme, leading to NO production, whereas mutant eNOS (S1179A) is resistant to phosphorylation and activation by Akt. Moreover, using adenovirus-mediated gene transfer, activated Akt increases basal NO release from endothelial cells, and activation-deficient Akt attenuates NO production stimulated by vascular endothelial growth factor. Thus, eNOS is a newly described Akt substrate linking signal transduction by Akt to the release of the gaseous second messenger NO.
Fulton
D
D
Department of Pharmacology, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Connecticut 06536, USA.
Gratton
J P
JP
McCabe
T J
TJ
Fontana
J
J
Fujio
Y
Y
Walsh
K
K
Franke
T F
TF
Papapetropoulos
A
A
Sessa
W C
WC
D000818:Animals; D019556:COS Cells; D002417:Cattle; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D051058:Oncogene Protein v-akt; D010766:Phosphorylation; D051381:Rats; D015682:Retroviridae Proteins, Oncogenic / Q000378:metabolism*; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection
D015682:Retroviridae Proteins, Oncogenic; D009569:Nitric Oxide; D012694:Serine; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat; D051058:Oncogene Protein v-akt