title: Differential binding characteristics and cellular inhibition by soluble VEGF receptors 1 and 2.
abstract: The FLT-1 and KDR genes encode transmembrane tyrosine kinases which function as high-affinity receptors for vascular endothelial growth factor (VEGF). We have used the baculovirus system to express the extracellular parts of the FLT-1 receptor and KDR receptor in soluble form (sFLT-1 and sKDR), for in vitro binding and competition assays. Here, we show that the binding of VEGF165 to sKDR but not sFLT-1 is dependent on heparin, regardless of whether VEGF165 or sKDR is immobilized. Further, only sFLT-1 acts as a receptor antagonist in solution and sKDR can neither compete with the binding of VEGF165 to human endothelial cells carrying both receptors nor block VEGF165 induced mitogenicity. Soluble KDR only partially inhibits cell migration even at high concentrations, in contrast to sFLT which can almost completely block (82%) VEGF-induced cell proliferation and migration. Taken together these results show that the two soluble VEGF receptor proteins, sFLT-1 and sKDR, despite binding the same ligand, behave very differently when immobilized with regard to their dependence on heparin for VEGF binding. In solution their respective ability to function as receptor antagonists is also strikingly different, possibly a reflection of their different dependency on heparin.
authors.affiliation: Department of Gene Regulation, GBF, Braunschweig, Germany.
authors.forename: W
authors.initials: W
authors.lastname: Roeckl
authors.forename: D
authors.initials: D
authors.lastname: Hecht
authors.forename: H
authors.initials: H
authors.lastname: Sztajer
authors.forename: J
authors.initials: J
authors.lastname: Waltenberger
authors.forename: A
authors.initials: A
authors.lastname: Yayon
authors.forename: H A
authors.initials: HA
authors.lastname: Weich
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D019281:Dimerization; D016228:Endothelial Growth Factors / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000494:pharmacology; D015870:Gene Expression / Q000235:genetics; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008024:Ligands; D008222:Lymphokines / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008934:Mitogens / Q000037:antagonists & inhibitors / Q000378:metabolism; D010969:Plastics; D011485:Protein Binding / Q000187:drug effects; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017978:Receptors, Growth Factor / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D040262:Receptors, Vascular Endothelial Growth Factor; D012995:Solubility; D042461:Vascular Endothelial Growth Factor A; D040281:Vascular Endothelial Growth Factor Receptor-1; D042442:Vascular Endothelial Growth Factors
chemical_list: D003432:Cross-Linking Reagents; D016228:Endothelial Growth Factors; D005343:Fibrinolytic Agents; D008024:Ligands; D008222:Lymphokines; D008934:Mitogens; D010969:Plastics; D011518:Proto-Oncogene Proteins; D017978:Receptors, Growth Factor; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D006493:Heparin; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor; D040281:Vascular Endothelial Growth Factor Receptor-1