Biotin binders selected from a random peptide library expressed on phage.
Recombinant biotin-binding phages were affinity-selected from a random peptide library expressed on the surface of filamentous phage. Phage binding to biotinylated proteins was half-maximally inhibited by micromolar concentrations of a monobiotinylated molecule. Sequencing of the peptide inserts of selected phages led to the identification of a previously unknown biotin-binding motif, CXWXPPF(K or R)XXC. A synthetic peptide containing this sequence motif inhibited streptavidin binding to biotinylated BSA with an IC50 of 50 microM. This compound represents the shortest non-avidin biotin-binding peptide identified to date. Our results illustrate that phage display technology can be used to identify novel ligands for a small non-proteinaceous molecule.
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Istituto di Ricerche di Biologia Molecolare P. Angeletti, Pomezia Roma, Italy.
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D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics* / Q000276:immunology; D001710:Biotin / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010455:Peptides; D001710:Biotin