Glycosylation of vascular endothelial growth factor is not required for its mitogenic activity.
We have stably expressed the cDNA encoding the 165 amino-acid long form of human vascular endothelial growth factor (VEGF) in BHK-21 cells. VEGF was partially purified from the conditioned medium of transfected cells using heparin-sepharose affinity chromatography. The partially purified VEGF was mitogenic for various types of endothelial cells and inhibited the binding of pure [125I]VEGF to its receptors. Western blot analysis, using anti-VEGF antibodies, revealed a 47 kDa VEGF homodimer in the partially purified VEGF fraction. Preincubation of the transfected cells with the N-glycosylation inhibitor tunicamycin resulted in the conversion of the 47 kDa VEGF homodimer into a smaller, deglycosylated form of 42 kDa. Partially purified preparations of the deglycosylated VEGF displayed a mitogenic activity that was similar to that of the glycosylated form and efficiently inhibited the binding of native [125I]VEGF to the VEGF receptors of bovine aortic arch derived endothelial cells.
Peretz
D
D
Department of Biology, Israel Institute of Technology, Technion City, Haifa.
Gitay-Goren
H
H
Safran
M
M
Kimmel
N
N
Gospodarowicz
D
D
Neufeld
G
G
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D016228:Endothelial Growth Factors / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D006031:Glycosylation; D006801:Humans; D008222:Lymphokines / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D008934:Mitogens / Q000235:genetics; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D015183:Restriction Mapping; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D008222:Lymphokines; D008934:Mitogens; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D004247:DNA