Lactoferrin-derived Peptides Active towards Influenza: Identification of Three Potent Tetrapeptide Inhibitors

Bovine lactoferrin is a biglobular multifunctional iron binding glycoprotein that plays an important role in innate immunity against infections. We have previously demonstrated that selected peptides from bovine lactoferrin C-lobe are able to prevent both Influenza virus hemagglutination and cell infection. To deeper investigate the ability of lactoferrin derived peptides to inhibit Influenza virus infection, in this study we identified new bovine lactoferrin C-lobe derived sequences and corresponding synthetic peptides were synthesized and assayed to check their ability to prevent viral hemagglutination and infection. We identified three tetrapeptides endowed with broad anti-Influenza activity and able to inhibit viral infection in a concentration range femto- to picomolar. Our data indicate that these peptides may constitute a non-toxic tool for potential applications as anti-Influenza therapeutics.

[1]  Jerson L. Silva,et al.  Inhibition of Mayaro virus infection by bovine lactoferrin. , 2014, Virology.

[2]  M. Ammendolia,et al.  Bovine lactoferrin-derived peptides as novel broad-spectrum inhibitors of influenza virus , 2012, Pathogens and global health.

[3]  K. Wüthrich.  NMR of proteins and nucleic acids , 1988 .

[4]  N. Zinder,et al.  The growth of an RNA bacteriophage: the role of DNA synthesis. , 1962, Virology.

[5]  M. Viljoen,et al.  Lactoferrin: a general review. , 1995, Haematologica.

[6]  E. L. Crossley.  A GENERAL REVIEW , 1954 .

[7]  Andrew L Hopkins,et al.  Fragment screening by surface plasmon resonance. , 2010, ACS medicinal chemistry letters.

[8]  M. Ammendolia,et al.  Bovine lactoferrin: involvement of metal saturation and carbohydrates in the inhibition of influenza virus infection. , 2012, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[9]  M. Ammendolia,et al.  Bovine lactoferrin prevents the entry and intercellular spread of herpes simplex virus type 1 in Green Monkey Kidney cells. , 2007, Antiviral research.

[10]  Shuwen Liu,et al.  Investigational hemagglutinin-targeted influenza virus inhibitors , 2017, Expert opinion on investigational drugs.

[11]  A. Osterhaus,et al.  Comparison of RNA hybridization, hemagglutination assay, titration of infectious virus and immunofluorescence as methods for monitoring influenza virus replication in vitro. , 1998, Journal of virological methods.

[12]  K. Wüthrich,et al.  Torsion angle dynamics for NMR structure calculation with the new program DYANA. , 1997, Journal of molecular biology.

[13]  S. Tanaka,et al.  Inhibition with lactoferrin of in vitro infection with human herpes virus. , 1994, Japanese journal of medical science & biology.

[14]  Richard R. Ernst,et al.  Elucidation of cross relaxation in liquids by two-dimensional N.M.R. spectroscopy , 1980 .

[15]  Gira Bhabha,et al.  Antibody Recognition of a Highly Conserved Influenza Virus Epitope , 2009, Science.

[16]  D. G. Davis,et al.  Practical aspects of two-dimensional transverse NOE spectroscopy , 1985 .

[17]  F. Belardelli,et al.  Antiviral activity of lactoferrin. , 1998, Advances in experimental medicine and biology.

[18]  S. Kaneko,et al.  Hepatitis C virus envelope proteins bind lactoferrin , 1997, Journal of virology.

[19]  E. De Clercq,et al.  Antiviral effects of plasma and milk proteins: lactoferrin shows potent activity against both human immunodeficiency virus and human cytomegalovirus replication in vitro. , 1995, The Journal of infectious diseases.

[20]  E. Novellino,et al.  Development and Identification of a Novel Anti-HIV-1 Peptide Derived by Modification of the N-Terminal Domain of HIV-1 Integrase , 2016, Front. Microbiol..

[21]  N. Kato,et al.  Lactoferrin inhibits hepatitis B virus infection in cultured human hepatocytes. , 2002, Hepatology research : the official journal of the Japan Society of Hepatology.

[22]  H. Tsutsumi,et al.  Lactoferrin and surfactant protein A exhibit distinct binding specificity to F protein and differently modulate respiratory syncytial virus infection , 2003, European journal of immunology.

[23]  PietrantoniAgostina,et al.  Bovine lactoferrin: involvement of metal saturation and carbohydrates in the inhibition of influenza virus infection11This article is part of a Special Issue entitled Lactoferrin and has undergone the Journal's usual peer review process. , 2012 .

[24]  B. Dewald,et al.  A novel type of cytoplasmic granule in bovine neutrophils , 1983, The Journal of cell biology.

[25]  R. Sheppard,et al.  Solid phase peptide synthesis : a practical approach , 1989 .

[26]  T. F. Smith,et al.  Replication and plaque assay of influenza virus in an established line of canine kidney cells. , 1968, Applied microbiology.

[27]  Ad Bax,et al.  MLEV-17-based two-dimensional homonuclear magnetization transfer spectroscopy , 1985 .

[28]  D. Colavizza,et al.  Properties of the iron-binding site of the N-terminal lobe of human and bovine lactotransferrins. Importance of the glycan moiety and of the non-covalent interactions between the N- and C-terminal lobes in the stability of the iron-binding site. , 1990, The Biochemical journal.

[29]  J. Skehel,et al.  The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. , 1987, Annual review of biochemistry.

[30]  Conrad C. Huang,et al.  UCSF Chimera—A visualization system for exploratory research and analysis , 2004, J. Comput. Chem..

[31]  G. Bifulco,et al.  Molecular insights into azumamide e histone deacetylases inhibitory activity. , 2007, Journal of the American Chemical Society.

[32]  F. Superti,et al.  Inhibition of herpes simplex virus infection by lactoferrin is dependent on interference with the virus binding to glycosaminoglycans. , 2004, Virology.

[33]  M. Harmsen,et al.  Antiviral activities of lactoferrin. , 2001, Antiviral research.

[34]  G. Bifulco,et al.  Synthesis and biological activity of cyclotetrapeptide analogues of the natural HDAC inhibitor FR235222. , 2010, Bioorganic & medicinal chemistry.

[35]  G. Bifulco,et al.  Molecular modeling studies toward the structural optimization of new cyclopeptide-based HDAC inhibitors modeled on the natural product FR235222. , 2008, Bioorganic & medicinal chemistry.

[36]  L. Vorland.  Lactoferrin: A multifunctional glycoprotein , 1999, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[37]  J. Thornton,et al.  PROMOTIF—A program to identify and analyze structural motifs in proteins , 1996, Protein science : a publication of the Protein Society.

[38]  V. Adam,et al.  Perspective of Use of Antiviral Peptides against Influenza Virus , 2015, Viruses.

[39]  J. Skehel,et al.  H1 and H7 influenza haemagglutinin structures extend a structural classification of haemagglutinin subtypes. , 2004, Virology.

[40]  Mariangela Agamennone,et al.  Identification of small molecules acting against H1N1 influenza A virus. , 2016, Virology.

[41]  J. Reynes,et al.  The fusion inhibitor enfuvirtide in recent antiretroviral strategies , 2009, Current opinion in HIV and AIDS.

[42]  Wei-June Chen,et al.  Bovine lactoferrin inhibits Japanese encephalitis virus by binding to heparan sulfate and receptor for low density lipoprotein. , 2008, Virology.

[43]  M. Ammendolia,et al.  Glycosaminoglycans are not indispensable for the anti-herpes simplex virus type 2 activity of lactoferrin. , 2009, Biochimie.

[44]  Steven J. Gamblin,et al.  Influenza Hemagglutinin and Neuraminidase Membrane Glycoproteins , 2010, The Journal of Biological Chemistry.

[45]  J. Thornton,et al.  AQUA and PROCHECK-NMR: Programs for checking the quality of protein structures solved by NMR , 1996, Journal of biomolecular NMR.

[46]  Richard R. Ernst,et al.  Multiple quantum filters for elucidating NMR coupling networks , 1982 .

[47]  M. G. Ciufolini,et al.  Bovine Lactoferrin Inhibits Toscana Virus Infection by Binding to Heparan Sulphate , 2015, Viruses.

[48]  T. Mizutani,et al.  In vitro antiviral activity of lactoferrin and ribavirin upon hantavirus , 2000, Archives of Virology.

[49]  M. Harmsen,et al.  Inhibition of cytomegalovirus infection by lactoferrin in vitro and in vivo. , 2004, Antiviral research.

[50]  I. Donatelli,et al.  Bovine lactoferrin inhibits Influenza A virus induced programmed cell death in vitro , 2010, BioMetals.

[51]  K. Wüthrich,et al.  Application of phase sensitive two-dimensional correlated spectroscopy (COSY) for measurements of 1H-1H spin-spin coupling constants in proteins. , 1983, Biochemical and biophysical research communications.

[52]  C. Groom,et al.  Three-dimensional structure of diferric bovine lactoferrin at 2.8 A resolution. , 1997, Journal of molecular biology.