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Screening of Peptide Ligands for Pyrroloquinoline Quinone Glucose Dehydrogenase Using Antagonistic Template-Based Biopanning
Department of Biotechnology, Graduate School of Engineering, Tokyo University of Agriculture & Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan
* Author to whom correspondence should be addressed.
Received: 31 July 2013; in revised form: 31 October 2013 / Accepted: 11 November 2013 / Published: 25 November 2013
Abstract: We have developed a novel method, antagonistic template-based biopanning, for screening peptide ligands specifically recognizing local tertiary protein structures. We chose water-soluble pyrroloquinoline quinone (PQQ) glucose dehydrogenase (GDH-B) as a model enzyme for this screening. Two GDH-B mutants were constructed as antagonistic templates; these have some point mutations to induce disruption of local tertiary structures within the loop regions that are located at near glucose-binding pocket. Using phage display, we selected 12-mer peptides that specifically bound to wild-type GDH-B but not to the antagonistic templates. Consequently, a peptide ligand showing inhibitory activity against GDH-B was obtained. These results demonstrate that the antagonistic template-based biopanning is useful for screening peptide ligands recognizing the specific local tertiary structure of proteins.
Keywords: peptide ligand; phage display; site-directed; inhibitor
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MDPI and ACS Style
Abe, K.; Yoshida, W.; Terada, K.; Yagi-Ishii, Y.; Ferri, S.; Ikebukuro, K.; Sode, K. Screening of Peptide Ligands for Pyrroloquinoline Quinone Glucose Dehydrogenase Using Antagonistic Template-Based Biopanning. Int. J. Mol. Sci. 2013, 14, 23244-23256.
Abe K, Yoshida W, Terada K, Yagi-Ishii Y, Ferri S, Ikebukuro K, Sode K. Screening of Peptide Ligands for Pyrroloquinoline Quinone Glucose Dehydrogenase Using Antagonistic Template-Based Biopanning. International Journal of Molecular Sciences. 2013; 14(12):23244-23256.
Abe, Koichi; Yoshida, Wataru; Terada, Kotaro; Yagi-Ishii, Yukiko; Ferri, Stefano; Ikebukuro, Kazunori; Sode, Koji. 2013. "Screening of Peptide Ligands for Pyrroloquinoline Quinone Glucose Dehydrogenase Using Antagonistic Template-Based Biopanning." Int. J. Mol. Sci. 14, no. 12: 23244-23256.