EDF-1 contributes to the regulation of nitric oxide release in VEGF-treated human endothelial cells

Marzia Leidi, Massimo Mariotti, Jeanette A M Maier

Research output: Contribution to journalArticle

Abstract

Vascular endothelial growth factor (VEGF) induces nitric oxide (NO) release by triggering multiple intracellular signals, among others the calcium/calmodulin pathway and the activation of Akt, events which induce endothelial NO synthase (eNOS) activity. Because Endothelial Differentiation-related Factor (EDF)-1 is a calmodulin binding protein and plays a role in modulating endothelial functions, we evaluated whether EDF-1 is implicated in the regulation of eNOS activity in VEGF-treated human endothelial cells. While VEGF does not modulate the total amounts of EDF-1, it promotes the dissociation of calmodulin from EDF-1 which correlates with the increase of calmodulin bound to eNOS and the induction of NO release. To better characterize the contribution of EDF-1 to the regulation of VEGF-induced NO release, we stably silenced EDF-1 in endothelial cells. We here show that endothelial cells silencing EDF-1 produce more NO than controls and do not increase NO release in response to VEGF. The insensitivity to VEGF results from the incapability of cells silencing EDF-1 to phosphorylate eNOS Ser 1177, even though Akt is activated. Interestingly, okadaic acid, a pharmacologic inhibitor of the serine/threonine phosphatase PP2A, which preferentially dephosphorylates eNOS Ser 1177, restores NO release and eNOS Ser 1177 phosphorylation in cells silencing EDF-1. Our results suggest EDF-1 as a novel contributor to the complex regulation of eNOS activity in human endothelial cells.

Original languageEnglish
Pages (from-to)654-660
Number of pages7
JournalEuropean Journal of Cell Biology
Volume89
Issue number9
DOIs
Publication statusPublished - Sep 2010

Fingerprint

Nitric Oxide Synthase
Nitric Oxide
Endothelial Cells
Vascular Endothelial Growth Factor A
Calmodulin
Calmodulin-Binding Proteins
Okadaic Acid
Nitric Oxide Synthase Type III
Phosphoprotein Phosphatases
Human Activities
human VEGFA protein
Phosphorylation
Calcium

Keywords

  • EDF-1
  • Human endothelial cells
  • Nitric oxide
  • VEGF

ASJC Scopus subject areas

  • Cell Biology
  • Histology
  • Pathology and Forensic Medicine

Cite this

EDF-1 contributes to the regulation of nitric oxide release in VEGF-treated human endothelial cells. / Leidi, Marzia; Mariotti, Massimo; Maier, Jeanette A M.

In: European Journal of Cell Biology, Vol. 89, No. 9, 09.2010, p. 654-660.

Research output: Contribution to journalArticle

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