Novel short peptides isolated from phage display library inhibit vascular endothelial growth factor activity.
Signal transduction through the vascular endothelial growth factor (VEGF)-VEGF receptor (VEGFR) pathway has a pivotal importance in angiogenesis, and has therefore become a prime target in antitumor therapy. In search for peptides antagonizing VEGF binding to its receptors, we screened a random heptamer library displayed on phage for peptides that bind the whole VEGF165 molecule and inhibit VEGF dependent human umbilical vein endothelial cell (HUVEC) proliferation. Two selected peptides with sequences WHLPFKC and WHKPFRF were synthesized. Biacore and matrix-assisted laser desorption/ionization timeof- flight mass spectrometry analysis indicated that these peptides bind the VEGF homodimer in a concentration- dependent manner, with micromolar affinity, and with a 2:1 peptide:VEGF stoichiometry. They inhibited HUVEC proliferation in vitro by 77 and 55%, respectively. Taken together, our results indicate that these peptides could be potent inhibitors of angiogenesis. Furthermore, we show that the peptide- VEGF binding properties can be quantified, a prerequisite for the further optimization of binders.
Erdag
Berrin
B
TUBITAK Research Institute for Genetic Engineering and Biotechnology, P.O.Box 21, 41470 Gebze, Kocaeli, Turkey. berrin@gmbae.tubitak.gov.tr
Balcioglu
Koray B
KB
Kumbasar
Asli
A
Celikbicak
Omur
O
Zeder-Lutz
Gabrielle
G
Altschuh
Danièle
D
Salih
Bekir
B
Baysal
Kemal
K
D000595:Amino Acid Sequence; D020533:Angiogenesis Inhibitors / Q000737:chemistry / Q000494:pharmacology; D019149:Bioreactors; D001709:Biotechnology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology*; D019151:Peptide Library; D011485:Protein Binding; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000494:pharmacology
D020533:Angiogenesis Inhibitors; D009842:Oligopeptides; D019151:Peptide Library; D011994:Recombinant Proteins; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A