ORIGINAL ARTICLE  

Identification of Antibiotic Clarithromycin Binding Peptide Displayed by T7 Phage Particles

Tetsuro Morimura, Naoko Noda, Yasutaro Kato, Tetsuaki Watanabe, Takeki Saitoh, Takayuki Yamazaki, Keiichi Takada, Satoko Aoki, Keisuke Ohta, Masahiko Ohshige, Kengo Sakaguchi, Fumio Sugawara

Received: May 15, 2006 / Accepted: September 25, 2006
© Japan Antibiotics Research Association

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 2.1×10-3 M. The dissociation constants of other macrolide antibiotics, erythromycin, roxithromycin, azithromycin and josamycin were also determined to be 5.4×10-3 M, 6.2×10-5 M, 1.1 M and 3.4×10-2 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.

Keywords phage display, macrolide antibiotics, clarithromycin, surface plasmon resonance


F. Sugawara (Corresponding author), T. Morimura, N. Noda, Y. Kato, T. Watanabe, T. Saitoh, T. Yamazaki, K. Takada, S. Aoki, K. Ohta, M. Ohshige, K. Sakaguchi: Genome and Drug Research Center, Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba 278-8510, Japan,
E-mail: sugawara@rs.noda.tus.ac.jp