Biochemical and Biophysical Research Communications
Augmentation of vascular permeability of VEGF is enhanced by KDR-binding proteins
Section snippets
Materials and methods
Materials. Snake venom proteins, vammin and KDR-bp, were purified from crude venom of the snakes as described previously [19], [20]. Recombinant human VEGF165 and TIMP-3 were purchased from PeproTech (Rocky Hill, NJ, USA) and R&D Systems (Minneapolis, MN, USA), respectively. Nω-nitro-l-arginine (l-NNA) was obtained from Sigma (St. Louis, MO, USA).
Miles assay. Vascular permeability was measured by using Miles assay [22]. After anesthesia by ketamine, guinea pigs (400 g) were shaved on the back
VEGF-F is a highly potent enhancer of vascular permeability
To clarify the correlation between vascular permeability-enhancing effects and receptor-binding of VEGFs, we first examined the effect of vammin, a snake venom-derived KDR-specific VEGF-F [19], on vascular leakage. Permeability-enhancing activity was measured by the Miles dye leakage assay [22]. While both vammin and VEGF165 similarly provoked dye leakage when injected intradermally at low concentrations (<10−8 M) vammin was a more potent enhancer of vascular leakage than VEGF165 at higher
Acknowledgments
We thank Dr. Craig M. Jackson for helping to revise the language of the manuscript. This work was supported in part by Scientific Research Grants-in-Aid from the Ministry of Education, Science, and Culture of Japan (T.M.), Mitsubishi Pharma Research Foundation (Y.Y.), and President’s Special Research Grant from Meiji Pharmaceutical University (Y.Y.).
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