VEGF(121), 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, VEGF(121) 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 VEGF(121) and VEGF(165) bound to the VEGF receptors of these cells, Unexpectedly, heparin inhibited the binding of VEGF(121) as well as the binding of VEGF(165) to the VEGF receptors of the melanoma cells, Digestion of the cells with heparinase also inhibited the binding of both VEGF variants, The VEGF(165) 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 VEGF(121) 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 VEGF(121) 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.