VEGF121, a vascular endothelial growth factor (VEGF) isoform lacking heparin binding ability, requires cell-surface heparan sulfates for efficient binding to the VEGF receptors of human melanoma cells.
Four vascular endothelial growth factor (VEGF) splice variants containing 121, 165, 189, and 206 amino acids are produced from a single human gene as a result of alternative splicing. VEGF121 is not a heparin-binding protein, while the other VEGF species possess heparin binding ability. YU-ZAZ6 human melanoma cells expressed the mRNA encoding the VEGF receptor flt-1, but not the mRNA encoding the VEGF receptor KDR/flk-1. Both VEGF121 and VEGF165 bound to the VEGF receptors of these cells. Unexpectedly, heparin inhibited the binding of VEGF121 as well as the binding of VEGF165 to the VEGF receptors of the melanoma cells. Digestion of the cells with heparinase also inhibited the binding of both VEGF variants. The VEGF165 binding ability of heparinase-digested cells could be partially restored by the addition of exogenous heparin to the binding reaction. In contrast, the addition of heparin to heparinase-digested cells did not restore VEGF121 binding. These results suggest that cell-surface heparan sulfates may regulate the binding ability of the VEGF receptors of the melanoma cells. They also indicate that heparin is not able to fully substitute for cell surface-associated heparan sulfates since VEGF121 binding to the VEGF receptors of heparinase-treated cells is not restored by heparin. These data suggest that changes in the composition of cell-surface heparin-like molecules may differentially affect the interaction of various VEGF isoforms with VEGF receptors.
Cohen
T
T
Department of Biology, Technion-Israel Institute of Technology, Haifa.
Gitay-Goren
H
H
Sharon
R
R
Shibuya
M
M
Halaban
R
R
Levi
B Z
BZ
Neufeld
G
G
D017398:Alternative Splicing; D001665:Binding Sites; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003432:Cross-Linking Reagents; D016228:Endothelial Growth Factors / Q000096:biosynthesis / Q000378:metabolism* / Q000494:pharmacology*; D015870:Gene Expression; D014644:Genetic Variation; D006493:Heparin / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008222:Lymphokines / Q000096:biosynthesis / Q000378:metabolism* / Q000494:pharmacology*; D008545:Melanoma / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism*; D017978:Receptors, Growth Factor / Q000096:biosynthesis / Q000378:metabolism*; D011975:Receptors, Mitogen / Q000378:metabolism; D040262:Receptors, Vascular Endothelial Growth Factor; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D003432:Cross-Linking Reagents; D016228:Endothelial Growth Factors; D006493:Heparin; D006497:Heparan Sulfate; D008222:Lymphokines; D012333:RNA, Messenger; D020794:Receptor Protein-Tyrosine Kinases; D017978:Receptors, Growth Factor; D011975:Receptors, Mitogen; D040262:Receptors, Vascular Endothelial Growth Factor; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; C467484:VEGFA protein, human