title: Identification of antibiotic clarithromycin binding peptide displayed by T7 phage particles.
abstract: Peptide libraries displayed by T7 phage, which contain random cDNA fragments insets, were screened for their ability to bind to a biotinylated derivative of clarithromycin. Phage particles bound to an immobilized derivative of the antibiotic were isolated and the inserted cDNA was amplified and sequenced. A common selected peptide sequence, composed of 19 amino acids, was obtained and a synthetic peptide with this sequence was produced. Surface plasmon resonance experiments showed that the synthetic peptide immobilized on a sensor chip bound to clarithromycin and the dissociation constant was determined to be 2.1 x 10(-3) M. The dissociation constants of other macrolide antibiotics, erythromycin, roxithromycin, azithromycin and josamycin were also determined to be 5.4 x 10(-3) M, 6.2 x 10(-5) M, 1.1 M and 3.4 x 10(-2) M, respectively. These results indicated that T7 phage display method might be useful to determine relatively weak interactions between small molecule drugs and the selected peptides which could represent a possible binding site conserved in binding proteins.
authors.affiliation: Genome and Drug Research Center, Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba 278-8510, Japan.
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Morimura
authors.forename: Naoko
authors.initials: N
authors.lastname: Noda
authors.forename: Yasutaro
authors.initials: Y
authors.lastname: Kato
authors.forename: Tetsuaki
authors.initials: T
authors.lastname: Watanabe
authors.forename: Takeki
authors.initials: T
authors.lastname: Saitoh
authors.forename: Takayuki
authors.initials: T
authors.lastname: Yamazaki
authors.forename: Keiichi
authors.initials: K
authors.lastname: Takada
authors.forename: Satoko
authors.initials: S
authors.lastname: Aoki
authors.forename: Keisuke
authors.initials: K
authors.lastname: Ohta
authors.forename: Masahiko
authors.initials: M
authors.lastname: Ohshige
authors.forename: Kengo
authors.initials: K
authors.lastname: Sakaguchi
authors.forename: Fumio
authors.initials: F
authors.lastname: Sugawara
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D017123:Bacteriophage T7 / Q000235:genetics*; D019921:Biotinylation; D017291:Clarithromycin / Q000031:analogs & derivatives / Q000378:metabolism*; D015723:Gene Library; D008024:Ligands; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D020349:Surface Plasmon Resonance
chemical_list: D000900:Anti-Bacterial Agents; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D017291:Clarithromycin