title: A peptidomimetic antagonist of TNF-alpha-mediated cytotoxicity identified from a phage-displayed random peptide library.
abstract: Phage-displayed peptide libraries represent a vast collection of peptide sequences that can be used to identify novel therapeutic molecules. In this report, a 15-mer phage-displayed peptide library was used to identify potential TNF-alpha antagonists. After direct interaction of recombinant human TNF-alpha with the library, four randomly selected phage clones were shown to inhibit in a dose-dependent fashion both mouse and human TNF-alpha-induced cytotoxicity in vitro. DNA sequencing of the positive clones revealed a common amino acid sequence that does not bear any structural similarity to the known primary structures of the extracellular domains of either 55-kDa or 75-kDa TNF receptors. This sequence was synthesized, and the peptidomimotope was shown i) to bind to the recombinant human TNF-alpha using surface plasmon resonance (biosensor) technology and ii) to inhibit both recombinant mouse and human TNF-alpha-induced cytotoxicity in vitro in a dose-dependent fashion. These findings highlight the potential of phage-displayed random peptide libraries for the identification of novel low molecular antagonistic molecules that can block the biologic activities of TNF-alpha.
authors.affiliation: Department of Infections and Tropical Diseases, London School of Hygiene and Tropical Medicine, United Kingdom.
authors.forename: C L
authors.initials: CL
authors.lastname: Chirinos-Rojas
authors.forename: M W
authors.initials: MW
authors.lastname: Steward
authors.forename: C D
authors.initials: CD
authors.lastname: Partidos
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D002999:Clone Cells; D003602:Cytotoxicity, Immunologic / Q000187:drug effects*; D006801:Humans; D051379:Mice; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding / Q000276:immunology; D011994:Recombinant Proteins / Q000378:metabolism; D017421:Sequence Analysis; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D019151:Peptide Library; D010455:Peptides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha