β-Hairpin Peptide That Targets Vascular Endothelial Growth Factor (VEGF) Receptors

Donatella Diana,Claudio Arra,Lucia De Rosa,Carlo Pedone,Sara Auriemma,Rossella Di Stasi,Luca Domenico D'Andrea,Maria Caterina Turco,Roberto Fattorusso,Anna Basile

doi:10.1074/jbc.m111.257402

Abstract

VEGF receptors have been the target of intense research aimed to develop molecules able to inhibit or stimulate angiogenesis. Based on the x-ray structure of the complex placental growth factor-VEGF receptor 1(D2), we designed a VEGF receptor-binding peptide reproducing the placental growth factor β-hairpin region Gln(87)-Val(100) that is involved in receptor recognition. A conformational analysis showed that the designed peptide adopts the expected fold in pure water. Moreover, a combination of NMR interaction analysis and cell binding studies were used to demonstrate that the peptide targets VEGF receptors. The VEGF receptor 1(D2)-interacting residues were characterized at the molecular level, and they correspond to the residues recognizing the placental growth factor sequence Gln(87)-Val(100). Finally, the peptide biological activity was characterized in vitro and in vivo, and it showed a VEGF-like behavior. Indeed, the peptide activated VEGF-dependent intracellular pathways, induced endothelial cell proliferation and rescue from apoptosis, and promoted angiogenesis in vivo. This compound is one of the few peptides known with proangiogenic activity, which makes it a candidate for the development of a novel peptide-based drug for medical applications in therapeutic angiogenesis.

Highlights

Angiogenesis is a fundamental physiological process involving the formation of a blood vessel from a preexisting one
VEGFRs have been the target of intense research aimed at developing molecules able to inhibit or stimulate angiogenesis [28]
A few years ago, we undertook the structure-based design of small VEGFR peptide binders based on a structural mimic approach that consists of reproducing the three-dimensional arrangements of the ligand-interacting residues stabilizing the

Summary

Introduction

Angiogenesis is a fundamental physiological process involving the formation of a blood vessel from a preexisting one. A combination of NMR interaction analysis and cell binding studies were used to demonstrate that the peptide targets VEGF receptors. We report the design, conformational analysis in aqueous solution, receptor interaction studies by NMR, and biological properties of a 17-mer ␤-hairpin peptide, HPLW, based on the ␤-hairpin sequence 87–100 of PlGF.

Results

Conclusion