title: An antagonistic vascular endothelial growth factor (VEGF) variant inhibits VEGF-stimulated receptor autophosphorylation and proliferation of human endothelial cells.
abstract: Vascular endothelial growth factor (VEGF) is a potent mitogen with a unique specificity for endothelial cells and a key mediator of aberrant endothelial cell proliferation and vascular permeability in a variety of human pathological situations, such as tumor angiogenesis, diabetic retinopathy, rheumatoid arthritis, or psoriasis. VEGF is a symmetric homodimeric molecule with two receptor binding interfaces lying on each pole of the molecule. Herein we report on the construction and recombinant expression of an asymmetric heterodimeric VEGF variant with an intact receptor binding interface at one pole and a mutant receptor binding interface at the second pole of the dimer. This VEGF variant binds to VEGF receptors but fails to induce receptor activation. In competition experiments, the heterodimeric VEGF variant antagonizes VEGF-stimulated receptor autophosphorylation and proliferation of endothelial cells. A 15-fold excess of the heterodimer was sufficient to inhibit VEGF-stimulated endothelial cell proliferation by 50%, and a 100-fold excess resulted in an almost complete inhibition. By using a rational approach that is based on the structure of VEGF, we have shown the feasibility to construct a VEGF variant that acts as an VEGF antagonist.
authors.affiliation: Institute of Molecular Medicine, Tumor Biology Center, Freiburg, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Siemeister
authors.forename: M
authors.initials: M
authors.lastname: Schirner
authors.forename: P
authors.initials: P
authors.lastname: Reusch
authors.forename: B
authors.initials: B
authors.lastname: Barleon
authors.forename: D
authors.initials: D
authors.lastname: Marmé
authors.forename: G
authors.initials: G
authors.lastname: Martiny-Baron
mesh_terms: D001665:Binding Sites; D001667:Binding, Competitive; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003001:Cloning, Molecular; D019281:Dimerization; D016228:Endothelial Growth Factors / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014644:Genetic Variation; D006801:Humans; D007700:Kinetics; D008222:Lymphokines / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D017978:Receptors, Growth Factor / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D040262:Receptors, Vascular Endothelial Growth Factor; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000494:pharmacology; D014469:Umbilical Arteries; D014471:Umbilical Veins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D016228:Endothelial Growth Factors; D008222:Lymphokines; D017978:Receptors, Growth Factor; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor