A Recombinant Mutant Vascular Endothelial Growth Factor-C that Has Lost Vascular Endothelial Growth Factor Receptor-2 Binding, Activation, and Vascular Permeability Activities

• Published in: The Journal of biological chemistry 1998-03, Vol.273 (12), p.6599-6602
• Main Authors: Joukov, V, Kumar, V, Sorsa, T, Arighi, E, Weich, H, Saksela, O, Alitalo, K
• Format: Article
• Language: English
• Subjects: Animals; Capillary Permeability; Cattle; Cell Line; Endothelial Growth Factors - genetics; Endothelial Growth Factors - metabolism; Endothelial Growth Factors - physiology; Mutagenesis; Protein Binding; Receptor Protein-Tyrosine Kinases - metabolism; Receptors, Growth Factor - metabolism; Receptors, Vascular Endothelial Growth Factor; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Swine; Vascular Endothelial Growth Factor C
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.12.6599

The vascular endothelial growth factor (VEGF) and the VEGF-C promote growth of blood vessels and lymphatic vessels, respectively. VEGF activates the endothelial VEGF receptors (VEGFR) 1 and 2, and VEGF-C activates VEGFR-3 and VEGFR-2. Both VEGF and VEGF-C are also potent vascular permeability factors. Here we have analyzed the receptor binding and activating properties of several cysteine mutants of VEGF-C including those (Cys 156 and Cys 165 ), which in other platelet-derived growth factor/VEGF family members mediate interchain disulfide bonding. Surprisingly, we found that the recombinant mature VEGF-C in which Cys 156 was replaced by a Ser residue is a selective agonist of VEGFR-3. This mutant, designated ΔNΔC156S, binds and activates VEGFR-3 but neither binds VEGFR-2 nor activates its autophosphorylation or downstream signaling to the ERK/MAPK pathway. Unlike VEGF-C, ΔNΔC156S neither induces vascular permeability in vivo nor stimulates migration of bovine capillary endothelial cells in culture. These data point out the critical role of VEGFR-2-mediated signal transduction for the vascular permeability activity of VEGF-C and strongly suggest that the redundant biological effects of VEGF and VEGF-C depend on binding and activation of VEGFR-2. The ΔNΔC156S mutant may provide a valuable tool for the analysis of VEGF-C effects mediated selectively via VEGFR-3. The ability of ΔNΔC156S to form homodimers also emphasizes differences in the structural requirements for VEGF and VEGF-C dimerization.