title: Identification by the phage-display technique of peptides that bind to H7 flagellin of Escherichia coli.
abstract: The four peptides interacting with H7 flagellin of Escherichia coli were selected from a phage display library. The library was selected four times, and the interacting phage peptides were competitively eluted with H7 flagellin. An enzyme-linked immunosorbent assay (ELISA) showed that these peptides were reactive with the H7 flagellin in a dose-dependent manner. Among them, a D1 phage clone showed the highest binding affinity to the H7 flagellin. We synthesized the D1 peptide (LHIHRPTLSIQG) corresponding to the peptide-encoding region of the D1 phage clone. The synthetic peptide showed micro-molar affinity (EC(50) value=1.9 microM) for the H7 flagellin. Furthermore, this D1 peptide interacted more specifically with the H7 flagellin than with the other flagellins (H1, H5, H12, or H23) of E. coli. In situ hybridization clearly showed that the peptide only detected those cells harboring the H7 flagellin gene (fliC). The peptide may specifically bind to the H7 flagellin on the cell surface. These results suggest that the phage-display technique could be used as a tool for identifying peptides as an alternative to using a ligand as a diagnostic reagent in food products or in clinical testing.
authors.affiliation: Marine Biotechnology Institute, Heita, Kamaishi, Iwate 026-0001, Japan. teruhiko.ide@mbio.jp
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Ide
authors.forename: Sang-Ho
authors.initials: SH
authors.lastname: Baik
authors.forename: Takao
authors.initials: T
authors.lastname: Matsuba
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Harayama
mesh_terms: D000595:Amino Acid Sequence; D015373:Bacterial Typing Techniques / Q000379:methods; D001435:Bacteriophages; D004797:Enzyme-Linked Immunosorbent Assay; D019453:Escherichia coli O157 / Q000737:chemistry / Q000302:isolation & purification; D029968:Escherichia coli Proteins / Q000378:metabolism*; D005408:Flagellin / Q000378:metabolism*; D019151:Peptide Library; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D029968:Escherichia coli Proteins; D019151:Peptide Library; D010455:Peptides; D005408:Flagellin