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Regulation of endothelium-derived nitric oxide production by the protein kinase Akt

Nature volume 399, pages 597–601 (1999)Cite this article

An Erratum to this article was published on 19 August 1999

Abstract

Endothelial nitric oxide synthase (eNOS) is the nitric oxide synthase isoform responsible for maintaining systemic blood pressure, vascular remodelling and angiogenesis1,2,3,4. eNOS is phosphorylated in response to various forms of cellular stimulation5,6,7 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.

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Figure 1: Wild-type Akt, but not kinase-inactive Akt, increases NO release from cells expressing membrane-associated eNOS.
Figure 2: Phosphorylation of eNOS by active Akt in vitro and in vivo.
Figure 3: Evidence that Ser 1179 is functionally important for Akt-stimulated NO release.
Figure 4: Akt regulates the basal and stimulated production of NO in endothelial cells.

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Acknowledgements

We thank P. Martasek and B. S. Masters for recombinant eNOS; J. Liu for constructing and characterizing the myr-nNOS construct; D. Bredt and T. Billiar for NOS cDNAs; T.Zioncheck for human VEGF; Y. Chen for generating the eNOS S1179A E. Coli expression construct; and K. Williams, M. LoPresti and K. Stone in the Keck Facility for their help with identification of the eNOS phosphopeptides. This work was supported by grants from the NIH and American Heart Association. W.C.S. is an Established Investigator of the American Heart Association. J.P.G. is supported by fellowships from the Heart and Stroke Foundation of Canada and from FCAR.

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Authors and Affiliations

  1. Department of Pharmacology and Molecular Cardiobiology Program, Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, 06536, Connecticut, USA

    David Fulton, Jean-Philippe Gratton, Timothy J. McCabe, Jason Fontana, Andreas Papapetropoulos & William C. Sessa

  2. Cardiovascular Research, St. Elizabeth's Medical Center, Boston, 02135, Massachusetts, USA

    Yasushi Fujio & Kenneth Walsh

  3. Department of Pharmacology, Columbia University, New York, 10032, New York, USA

    Thomas F. Franke

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Correspondence to William C. Sessa.

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Fulton, D., Gratton, JP., McCabe, T. et al. Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature 399, 597–601 (1999). https://doi.org/10.1038/21218

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