IDENTIFICATION OF RECEPTOR-BINDING SITES BY COMPETITIVE PEPTIDE-MAPPING - PHAGE-T1, PHAGE-T5, AND PHAGE-PHI-80 AND COLICIN-M BIND TO THE GATING LOOP OF FHUA

Previously we proposed a transmembrane model of the FhuA receptor prot ein in the outer membrane of Escherichia coli, Removal of the largest loop at the cell surface converted the FhuA transport protein into an open channel and rendered cells resistant to the FhuA-specific phages T1, T5, and phi 80 and to colicin M, In the present study we employed acetylated hexapeptide amides covering the entire surface loop to inve stigate binding of the phages and of colicin M. Competitive peptide ma pping proved to be a powerful technique to uncover three ligand bindin g sites within a region of 34 amino acid residues, Hexapeptides derive d from three specific regions of the surface loop inhibited infection of cells by the phages and killing by colicin M. Two of these regions were common among all four FhuA ligands, Electron microscopy of phage T5 revealed that one inhibitory peptide triggered a strong conformatio nal change leading to the release of DNA from the phage head. These re sults suggest that the FhuA gating loop is the target for specific bin ding of phages T1, T5, and phi 80 and colicin M.