Fibroblast growth factor 2‐antagonist activity of a long‐pentraxin 3‐derived anti‐angiogenic pentapeptide

• Published in: Journal of cellular and molecular medicine 2010-08, Vol.14 (8), p.2109-2121
• Main Authors: Leali, Daria, Bianchi, Roberta, Bugatti, Antonella, Nicoli, Stefania, Mitola, Stefania, Ragona, Laura, Tomaselli, Simona, Gallo, Grazia, Catello, Sergio, Rivieccio, Vincenzo, Zetta, Lucia, Presta, Marco
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Angiogenesis; Angiogenesis Inhibitors - pharmacology; Animals; Antagonists; Antibodies; BIAcore; Binding sites; Binding Sites - genetics; Biological activity; C-Reactive Protein - chemistry; C-Reactive Protein - metabolism; Cell growth; Cell proliferation; Cell Proliferation - drug effects; Cell Transplantation - methods; Cellular biology; Chick Embryo; CHO Cells; chorioallantoic membrane; Cricetinae; Cricetulus; embryo; Endothelial cells; Endothelial Cells - metabolism; Endothelial Cells - transplantation; Female; FGF; Fibroblast growth factor 2; Fibroblast Growth Factor 2 - antagonists & inhibitors; Fibroblast Growth Factor 2 - genetics; Fibroblast Growth Factor 2 - metabolism; Fibroblast growth factor receptor 1; Fibroblast growth factors; Fibroblasts; Gene expression; Growth factors; heparan sulphate; Heparin; Humans; Hydrophobicity; Immunoglobulins; Kinases; Ligands; Magnetic Resonance Spectroscopy; Male; Mitogens; Mutation; Neovascularization, Physiologic - drug effects; NMR; Nuclear magnetic resonance; Oligopeptides - metabolism; Oligopeptides - pharmacology; Pattern recognition receptors; pentraxin; Pentraxins; Peptides; Protein Binding - drug effects; Proteins; Receptor, Fibroblast Growth Factor, Type 1 - metabolism; receptors; Rodents; Sensors; Serum Amyloid P-Component - chemistry; Serum Amyloid P-Component - metabolism; Synthetic peptides; Transplantation, Heterologous; tumour; Wound healing; Zebrafish
• ISSN: 1582-1838; 1582-4934
• DOI: 10.1111/j.1582-4934.2009.00855.x

Fibroblast growth factor‐2 (FGF2) plays a major role in angiogenesis. The pattern recognition receptor long‐pentraxin 3 (PTX3) inhibits the angiogenic activity of FGF2. To identify novel FGF2‐antagonistic peptide(s), four acetylated (Ac) synthetic peptides overlapping the FGF2‐binding region PTX3‐(97–110) were assessed for their FGF2‐binding capacity. Among them, the shortest pentapeptide Ac‐ARPCA‐NH2 (PTX3‐[100–104]) inhibits the interaction of FGF2 with PTX3 immobilized to a BIAcore sensorchip and suppresses FGF2‐dependent proliferation in endothelial cells, without affecting the activity of unrelated mitogens. Also, Ac‐ARPCA‐NH2 inhibits angiogenesis triggered by FGF2 or by tumorigenic FGF2‐overexpressing murine endothelial cells in chick and zebrafish embryos, respectively. Accordingly, the peptide hampers the binding of FGF2 to Chinese Hamster ovary cells overexpressing the tyrosine‐kinase FGF receptor‐1 (FGFR1) and to recombinant FGFR1 immobilized to a BIAcore sensorchip without affecting heparin interaction. In all the assays the mutated Ac‐ARPSA‐NH2 peptide was ineffective. In keeping with the observation that hydrophobic interactions dominate the interface between FGF2 and the FGF‐binding domain of the Ig‐like loop D2 of FGFR1, amino acid substitutions in Ac‐ARPCA‐NH2 and saturation transfer difference‐nuclear magnetic resonance analysis of its mode of interaction with FGF2 implicate the hydrophobic methyl groups of the pentapeptide in FGF2 binding. These results will provide the basis for the design of novel PTX3‐derived anti‐angiogenic FGF2 antagonists.