Identification of receptor binding sites by competitive peptide mapping: phages T1, T5, and phi 80 and colicin M bind to the gating loop of FhuA.
Previously we proposed a transmembrane model of the FhuA receptor protein 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 investigate binding of the phages and of colicin M. Competitive peptide mapping proved to be a powerful technique to uncover three ligand binding sites within a region of 34 amino acid residues. Hexapeptides derived 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 conformational change leading to the release of DNA from the phage head. These results suggest that the FhuA gating loop is the target for specific binding of phages T1, T5, and phi 80 and colicin M.
Universität Tübingen, Germany.
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H
Killmann
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G
Videnov
G
G
Jung
H
H
Schwarz
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V
Braun
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D001435:Bacteriophages / Q000187:drug effects / Q000378:metabolism*; D001665:Binding Sites; D003087:Colicins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology; D010449:Peptide Mapping; D011991:Receptors, Virus / Q000737:chemistry / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013604:T-Phages / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D003087:Colicins; D029968:Escherichia coli Proteins; C050900:FhuA protein, E coli; D010446:Peptide Fragments; D011991:Receptors, Virus