Distinct heparin binding sites on VEGF165 and its receptors revealed by their interaction with a non sulfated glycoaminoglycan (NaPaC).
We previously demonstrated that a non sulfated analogue of heparin, phenylacetate carboxymethyl benzylamide dextran (NaPaC) inhibited angiogenesis. Here, we observed that NaPaC inhibited the VEGF165 binding to both VEGFR2 and NRP-1 and abolished VEGFR2 activity. Further, we explored the effects of NaPaC on VEGF165 interactions with its receptors, VEGFR2 and NRP-1, co-receptor of VEGFR2. Surface plasmon resonance and affinity gel electrophoresis showed that NaPaC interacted directly with VEGF165, VEGFR2 and NRP-1 but not with heparin-independent factor such as VEGF121. NaPaC completely inhibited the heparin binding to VEGF165, NRP-1 and VEGFR2. We found that NaPaC bound to all three molecules, VEGF165, VEGFR2 and NRP-1, but was more effective in inhibiting heparin binding to VEGF165. These results suggested that heparin binding sites of VEGFR2 and NRP-1 were different from those of VEGF165.
Di Benedetto
Mélanie
M
CNRS-UMR7033, Equipe de pharmacologie, Université Paris 13, Bobigny, France. dibenedetto22@yahoo.fr
Starzec
Anna
A
Vassy
Roger
R
Perret
Gérard-Yves
GY
Crépin
Michel
M
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive / Q000187:drug effects; D015153:Blotting, Western; D002478:Cells, Cultured; D003911:Dextrans / Q000378:metabolism* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D039942:Neuropilin-1 / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013552:Swine; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism*
D003911:Dextrans; D020033:Protein Isoforms; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C476172:phenylacetate carboxymethyl benzylamide dextran; D039942:Neuropilin-1; D006493:Heparin; D040301:Vascular Endothelial Growth Factor Receptor-2