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

Leali D, Bianchi R, Bugatti A, Nicoli S, Mitola S, Ragona L, Tomaselli S, Gallo G, Catello S, Rivieccio V, Zetta L, Presta M - J. Cell. Mol. Med. (2010)

Bottom Line: In all the assays the mutated Ac-ARPSA-NH(2) 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-NH(2) 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.

View Article: PubMed Central - PubMed

Affiliation: Unit of General Pathology and Immunology, Department of Biomedical Sciences and Biotechnology, School of Medicine, University of Brescia, Brescia, Italy.

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Effect of Ac-ARPCA-NH2 on the biological activity of the FGF2/FGFR system. (A) GM 7373 cells were treated with FGF2 (0.55 nM), FGF1 (1.66 nM), FGF8b (1.66 nM), VEGF (0.7 nM), EGF (0.6 nM), DAG (15 μM), TPA (8.0 nM) or 10% FCS in the absence or in the presence of Ac-ARPCA-NH2 (66 μM). (B) CHO cells overexpressing FGFR1, FGFR2, FGFR3 or FGFR4 were treated with FGF2 (0.55 nM) in the absence or in the presence of Ac-ARPCA-NH2 (black bars) or Ac-ARPSA-NH2 (open bars) (both peptides at 300 μM). In both assays, cells were trypsinized and counted 24 hrs after the stimulus. Data (mean ± S.D. of triplicate observations) are expressed as percentage of cell proliferation measured in the absence of the peptide under test.
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fig01: Effect of Ac-ARPCA-NH2 on the biological activity of the FGF2/FGFR system. (A) GM 7373 cells were treated with FGF2 (0.55 nM), FGF1 (1.66 nM), FGF8b (1.66 nM), VEGF (0.7 nM), EGF (0.6 nM), DAG (15 μM), TPA (8.0 nM) or 10% FCS in the absence or in the presence of Ac-ARPCA-NH2 (66 μM). (B) CHO cells overexpressing FGFR1, FGFR2, FGFR3 or FGFR4 were treated with FGF2 (0.55 nM) in the absence or in the presence of Ac-ARPCA-NH2 (black bars) or Ac-ARPSA-NH2 (open bars) (both peptides at 300 μM). In both assays, cells were trypsinized and counted 24 hrs after the stimulus. Data (mean ± S.D. of triplicate observations) are expressed as percentage of cell proliferation measured in the absence of the peptide under test.

Mentions: PTX3 is able to inhibit the mitogenic activity exerted by FGF2 and by other members of the FGF family on endothelial cells, without affecting the activity of unrelated mitogens [28]. On this basis, ARPCA was assessed for its capacity to affect the proliferation of endothelial GM7373 cells exposed to different mitogenic stimuli. In these and following biological experiments, the inactive Ac-ARPSA-NH2 (hereafter referred to as ARPSA) was used as a negative control peptide. As shown in Fig. 1(A), ARPCA inhibits GM7373 cell proliferation triggered by FGF2, FGF8b and with a less efficiency by FGF1, whereas it does not affect the mitogenic activity of VEGF, EGF, DAG, TPA and serum. No inhibition was instead exerted by control peptide ARPSA on any mitogen. Moreover, in keeping with the capacity of FGF2 to interact with all four members of the FGFR family [47], ARPCA, but not ARPSA, inhibits the proliferation triggered by FGF2 in CHO cells stably transfected with the FGFR1, FGFR2, FGFR3 or FGFR4 isoforms [35] (Fig. 1B). These data demonstrate that, as for PTX3, the inhibitory activity of ARPCA is limited to the FGF/FGFR system and it is not due to a generic antiproliferative/toxic effect.


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

Leali D, Bianchi R, Bugatti A, Nicoli S, Mitola S, Ragona L, Tomaselli S, Gallo G, Catello S, Rivieccio V, Zetta L, Presta M - J. Cell. Mol. Med. (2010)

Effect of Ac-ARPCA-NH2 on the biological activity of the FGF2/FGFR system. (A) GM 7373 cells were treated with FGF2 (0.55 nM), FGF1 (1.66 nM), FGF8b (1.66 nM), VEGF (0.7 nM), EGF (0.6 nM), DAG (15 μM), TPA (8.0 nM) or 10% FCS in the absence or in the presence of Ac-ARPCA-NH2 (66 μM). (B) CHO cells overexpressing FGFR1, FGFR2, FGFR3 or FGFR4 were treated with FGF2 (0.55 nM) in the absence or in the presence of Ac-ARPCA-NH2 (black bars) or Ac-ARPSA-NH2 (open bars) (both peptides at 300 μM). In both assays, cells were trypsinized and counted 24 hrs after the stimulus. Data (mean ± S.D. of triplicate observations) are expressed as percentage of cell proliferation measured in the absence of the peptide under test.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3823002&req=5

fig01: Effect of Ac-ARPCA-NH2 on the biological activity of the FGF2/FGFR system. (A) GM 7373 cells were treated with FGF2 (0.55 nM), FGF1 (1.66 nM), FGF8b (1.66 nM), VEGF (0.7 nM), EGF (0.6 nM), DAG (15 μM), TPA (8.0 nM) or 10% FCS in the absence or in the presence of Ac-ARPCA-NH2 (66 μM). (B) CHO cells overexpressing FGFR1, FGFR2, FGFR3 or FGFR4 were treated with FGF2 (0.55 nM) in the absence or in the presence of Ac-ARPCA-NH2 (black bars) or Ac-ARPSA-NH2 (open bars) (both peptides at 300 μM). In both assays, cells were trypsinized and counted 24 hrs after the stimulus. Data (mean ± S.D. of triplicate observations) are expressed as percentage of cell proliferation measured in the absence of the peptide under test.
Mentions: PTX3 is able to inhibit the mitogenic activity exerted by FGF2 and by other members of the FGF family on endothelial cells, without affecting the activity of unrelated mitogens [28]. On this basis, ARPCA was assessed for its capacity to affect the proliferation of endothelial GM7373 cells exposed to different mitogenic stimuli. In these and following biological experiments, the inactive Ac-ARPSA-NH2 (hereafter referred to as ARPSA) was used as a negative control peptide. As shown in Fig. 1(A), ARPCA inhibits GM7373 cell proliferation triggered by FGF2, FGF8b and with a less efficiency by FGF1, whereas it does not affect the mitogenic activity of VEGF, EGF, DAG, TPA and serum. No inhibition was instead exerted by control peptide ARPSA on any mitogen. Moreover, in keeping with the capacity of FGF2 to interact with all four members of the FGFR family [47], ARPCA, but not ARPSA, inhibits the proliferation triggered by FGF2 in CHO cells stably transfected with the FGFR1, FGFR2, FGFR3 or FGFR4 isoforms [35] (Fig. 1B). These data demonstrate that, as for PTX3, the inhibitory activity of ARPCA is limited to the FGF/FGFR system and it is not due to a generic antiproliferative/toxic effect.

Bottom Line: In all the assays the mutated Ac-ARPSA-NH(2) 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-NH(2) 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.

View Article: PubMed Central - PubMed

Affiliation: Unit of General Pathology and Immunology, Department of Biomedical Sciences and Biotechnology, School of Medicine, University of Brescia, Brescia, Italy.

Show MeSH
Related in: MedlinePlus