Antiviral activity of ovotransferrin derived peptides

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Abstract

Ovotransferrin and lactoferrin are iron-binding proteins with antiviral and antibacterial activities related to natural immunity, showing marked sequence and structural homologies. The antiviral activity of two hen ovotransferrin fragments DQKDEYELL (hOtrf219–227) and KDLLFK (hOtrf269–301 and hOtrf633–638) towards Marek’s disease virus infection of chicken embryo fibroblasts is reported here. These fragments have sequence homology with two bovine lactoferrin fragments with antiviral activity towards herpes simplex virus, suggesting that these fragments could have a role for the exploitation of the antiviral activity of the intact proteins towards herpes viruses. NMR analysis showed that these peptides, chemically synthetized, did not possess any favourite conformation in solution, indicating that both the aminoacid sequence and the conformation they display in the intact protein are essential for the antiviral activity.

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Materials and methods

Proteins. Bovine milk lactoferrin was from Armor Proteins (Bretagne, France); human lactoferrin was from Sigma Chemical (St. Louis, MO); and hOtrf was purified by chicken white egg, as previously described [28]. In all cases, protein purity was checked by silver-stained SDS–PAGE. Protein concentration was determined by UV spectroscopy, assuming an extinction coefficient (280 nm, 1% solution) of 1.51 for lactoferrins [29] and 1.10 for ovotransferrin [28]. Before biological assays, all proteins

Results and discussion

Three bLf-derived peptides possess antiviral activity: lactoferricin B (bLf17–41) [22], [23], [24], fragments ADRDQYELL (bLf222–230) and EDLIWK (bLf264–269) [27], though their antiviral activity was much lower than that of the intact protein. The hOtrf fragments displaying sequence and/or structural homologies with antiviral bLf fragments are listed in Table 1. No fragment was identified in hOtrf having sequence homology with bLf fragment lactoferricin B (bLf17–41). On the contrary, two

Acknowledgments

The work was partially supported by Ministero Istruzione Universitàe Ricerca, Project COFIN 2002. Prof. P. Valenti (II University of Naples, Italy) and Prof. L. Seganti (University of Rome “La Sapienza”) are gratefully acknowledged for helpful discussions.

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    Abbreviations: bLf, bovine lactoferrin; hOtrf, hen ovotransferrin; MDV, Marek’s disease virus.

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