The DOI matches its Crossref record; 29 of the 29 references it cites were found in a registry or an index.
Identification of Regulatory Sites of Phosphorylation of the Bovine Endothelial Nitric-oxide Synthase at Serine 617 and Serine 635
Open accessAbstract
Endothelial nitric-oxide synthase (eNOS) is regulated by signaling pathways involving multiple sites of phosphorylation. The coordinated phosphorylation of eNOS at Ser(1179) and dephosphorylation at Thr(497) activates the enzyme, whereas inhibition results when Thr(497) is phosphorylated and Ser(1179) is dephosphorylated. We have identified two further phosphorylation sites, at Ser(617) and Ser(635), by phosphopeptide mapping and matrix-assisted laser desorption ionization time of flight mass spectrometry. Purified protein kinase A (PKA) phosphorylates both sites in purified eNOS, whereas purified Akt phosphorylates only Ser(617). In bovine aortic endothelial cells, bradykinin (BK), ATP, and vascular endothelial growth factor stimulate phosphorylation of both sites. BK-stimulated phosphorylation of Ser(617) is Ca(2+)-dependent and is partially inhibited by LY294002 and wortmannin, phosphatidylinositol 3-kinase inhibitors, suggesting signaling via Akt. BK-stimulated phosphorylation of Ser(635) is Ca(2+)-independent and is completely abolished by the PKA inhibitor, KT5720, suggesting signaling via PKA. Activation of PKA with isobutylmethylxanthine also causes Ser(635), but not Ser(617), phosphorylation. Mimicking phosphorylation at Ser(635) by Ser to Asp mutation results in a greater than 2-fold increase in activity of the purified protein, whereas mimicking phosphorylation at Ser(617) does not alter maximal activity but significantly increases Ca(2+)-calmodulin sensitivity. These data show that phosphorylation of both Ser(617) and Ser(635) regulates eNOS activity and contributes to the agonist-stimulated eNOS activation process.
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“The phosphorylation has been attributed to protein kinase C (PKC) (297, 646), and the residue is dephosphorylated by protein phosphatase 1 (PP1)”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [646]
In the citing paper
“Phosphorylation of Thr495 is a negative regulator of CaM binding to eNOS (297, 407, 646).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [646]
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AMP‐activated protein kinase phosphorylation of endothelial NO synthase
Chen, Z.-P., Mitchelhill, K. I., Michell, B. J., Stapleton, D., Rodríguez‐Crespo, I., Witters, L. A., Power, D. A., de Montellano, P. R. O., & Kemp, B. E. (1999) · FEBS Letters · doi:10.1016/s0014-5793(98)01705-0 · Cited by 839
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Self-citationMontagnani, M., Chen, H., Barr, V. A., & Quon, M. J. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m103702200 · Cited by 550
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- Fulton, 2001, J. Pharmacol. Exp. Ther., 299, 818
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Mlynek, A. (2012) · Univ. Duesseldorf: Duesseldorfer Dokumenten- und Publikationsserver
Lesyk, G. M. (2019) · ERA: Education and Research Archive (University of Alberta) · doi:10.7939/r3-pk4m-h082
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(2005) · CRC Press eBooks · doi:10.1201/9781420023312.bmatt
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Stimulation of eNOS-Ser617 Phosphorylation by Fluid Shear Stress in Endothelial Cells
Boo, Y. C. (2005) · Journal of Applied Biological Chemistry
Endothélium, inflammation et cognition : focus sur le BDNF
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Regulation of Translation by PKA Signaling Pathway
Yang, L., Hou, K., & Qi, H. (2026) · International Journal of Molecular Sciences · doi:10.3390/ijms27156789
“…a substrate of PKA, many metabolic enzymes and kinases are also found to be regulated by PKA, including endothelial nitric oxide synthase (eNOS), H + -ATPase and eukaryotic elongation factor-2 kinase (eEF2K) [66–68], indicating the close relationships among metabolism, translation and PKA activity.”
Boo, Y. C. (2005) · Journal of Applied Biological Chemistry
Metadata
- Authors
- Michell, B. J., Harris, M. B., Chen, Z.-P., Ju, H., Venema, V. J., Blackstone, M. A., Huang, W., Venema, R. C., & Kemp, B. E.
- Affiliations
- St Vincents Institute of Medical Research (AU)
- Center for Vascular Biology Research (US)
- Augusta University (US)
- Corresponding author
- Venema, Richard C.
- Kemp, Bruce E.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2002-11
- Volume
- 277
- Issue
- 44
- Pages
- 42344-42351
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2024185487
- Cited by
- 212
- References
- 29
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 2 · 2022: 1 · 2021: 7 · 2020: 4 · 2019: 5 · 2018: 4 · 2017: 8 · 2016: 9 · 2015: 5 · 2014: 13 · 2013: 7 · 2012: 15
- FWCI
- 9.39
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- American Heart Association
- National Institutes of Health
- Medical Research Council
- National Health and Medical Research Council
- Topics
- Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Health Sciences › Medicine › Surgery › Cardiovascular, Neuropeptides, and Oxidative Stress Research
- Keywords
- endothelial nitric oxide synthase, eNOS phosphorylation, protein kinase A, Akt, bradykinin signaling, phosphatidylinositol 3-kinase, enzyme activation, phosphopeptide mapping, bovine aortic endothelial cells
- MeSH terms
- Adenosine Triphosphate, Amino Acid Sequence, Animals, Bradykinin, Calcium, Calmodulin, Cattle, Humans, Lymphokines, Molecular Sequence Data, Phosphorylation, Proto-Oncogene Proteins, Serine, Endothelial Growth Factors, Protein Serine-Threonine Kinases, Cyclic AMP-Dependent Protein Kinases, Nitric Oxide Synthase, Intercellular Signaling Peptides and Proteins, Vascular Endothelial Growth Factors, Vascular Endothelial Growth Factor A, Proto-Oncogene Proteins c-akt, Nitric Oxide Synthase Type III
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