Biochemical and Biophysical Research Communications
VEGF-E activates endothelial nitric oxide synthase to induce angiogenesis via cGMP and PKG-independent pathways
Section snippets
Materials and methods
Reagents. Recombinant VEGF-E (orf virus strain D-1701) and VEGF-A were purchased from RELIATech (Brauschweig, Germany). Growth factor-reduced Matrigel was obtained from BD Biosciences (Cowley, UK). l-NNA (NG-NO2-l-arginine), ODQ (1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one), KT-5823 and U73122 were obtained from Calbiochem (Nottingham, UK). In vitro Angio kit was purchased from TCS Biologicals (Buckingham, UK). All other cell culture reagents and chemicals were obtained from Sigma Chemical
VEGF-E induces an in vitro angiogenic response
Immunoprecipitation and Western blot analysis were used to demonstrate that VEGF-E induces autophosphorylation of VEGFR-2 in PAEVEGFR-2 cells and HUVEC (Fig. 1A), but not in PAEVEGFR-1 cells (data not shown). VEGF-E was shown previously to induce the migration of PAEVEGFR-2, but not of PAEVEGFR-1[21]. In primary cultures, VEGF-E also increased endothelial cell migration (Fig. 1B). To determine whether selective VEGFR-2 stimulation alone can induce in vitro angiogenesis, the ability of VEGF-E to
Discussion
VEGF-E is likely to be a superior candidate for therapeutic angiogenesis as VEGF-A is associated with edema, hemorrhage and leukocyte recruitment [24]. This inflammatory response is not seen with VEGF-E overexpression [25]. This study provides the first evidence that activation of VEGFR-2 by VEGF-E induces phosphorylation of eNOS, stimulates the release of NO to promote angiogenesis in a cGMP- and PKG-independent manner. VEGF-E was identified as a VEGFR-2 specific viral homologue of VEGF-A
Acknowledgments
This work was supported by grants from the British Heart Foundation, the Wellcome Trust and the European Vascular Genomics Network, a network of excellence supported by the European Community’s Sixth Framework Programme for Research Priority 1 ‘Life Sciences, Genomics and Biotechnology for Health’ (contract LSHM-CT-2003-503254).
References (42)
- et al.
Role of eNOS in neovascularization: NO for endothelial progenitor cells
Trends Mol. Med.
(2004) - et al.
Vascular endothelial growth factor receptor-1 modulates vascular endothelial growth factor-mediated angiogenesis via nitric oxide
Am. J. Pathol.
(2001) - et al.
Vascular endothelial growth factor signals endothelial cell production of nitric oxide and prostacyclin through flk-1/KDR activation of c-Src
J. Biol. Chem.
(1999) - et al.
VEGF induces nuclear translocation of Flk-1/KDR, endothelial nitric oxide synthase, and caveolin-1 in vascular endothelial cells
Biochem. Biophys. Res. Commun.
(1999) - et al.
Parapoxviruses: potential alternative vectors for directing the immune response in permissive and non-permissive hosts
J. Biotechnol.
(1999) - et al.
A novel type of vascular endothelial growth factor, VEGF-E (NZ-7 VEGF), preferentially utilizes KDR/Flk-1 receptor and carries a potent mitotic activity without heparin-binding domain
J. Biol. Chem.
(1998) - et al.
Increased microvascular density and enhanced leukocyte rolling and adhesion in the skin of VEGF transgenic mice
J. Invest. Dermatol.
(1998) - et al.
VEGFR-2-specific ligand VEGF-E induces non-edematous hyper-vascularization in mice
Biochem. Biophys. Res. Commun.
(2003) - et al.
Different signal transduction properties of KDR and Flt1, two receptors for vascular endothelial growth factor
J. Biol. Chem.
(1994) - et al.
FGF-2, NGF and IGF-1, but not BDNF, utilize a nitric oxide pathway to signal neurotrophic and neuroprotective effects against alcohol toxicity in cerebellar granule cell cultures
Brain Res. Dev. Brain Res.
(2003)
Nitric oxide and cGMP activate the Ras-MAP kinase pathway-stimulating protein tyrosine phosphorylation in rabbit aortic endothelial cells
Free Radic. Biol. Med.
Nitric oxide inhibits proliferation of human endothelial cells via a mechanism independent of cGMP
Atherosclerosis
YC-1 inhibits proliferation of human vascular endothelial cells through a cyclic GMP-independent pathway
Biochem. Pharmacol.
G protein-coupled endothelial receptor for atypical cannabinoid ligands modulates a Ca2+-dependent K+ current
J. Biol. Chem.
Regulation of protein tyrosine phosphatase 1B in intact cells by S-nitrosothiols
Arch. Biochem. Biophys.
Nitric oxide reversibly inhibits seven members of the caspase family via S-nitrosylation
Biochem. Biophys. Res. Commun.
Nipradilol inhibits apoptosis by preventing the activation of caspase-3 via S-nitrosylation and the cGMP-dependent pathway
Eur. J. Pharmacol.
Presentation of nitric oxide regulates monocyte survival through effects on caspase-9 and caspase-3 activation
J. Biol. Chem.
Hypertension in mice lacking the gene for endothelial nitric oxide synthase
Nature
Vascular endothelial cells synthesize nitric oxide from l-arginine
Nature
Purification and characterization of particulate endothelium-derived relaxing factor synthase from cultured and native bovine aortic endothelial cells
Proc. Natl. Acad. Sci. USA
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These authors contributed equally to this work.