title: Screening of peptide ligands for pyrroloquinoline quinone glucose dehydrogenase using antagonistic template-based biopanning.
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.
authors.affiliation: Department of Biotechnology, Graduate School of Engineering, Tokyo University of Agriculture & Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan. sode@cc.tuat.ac.jp.
authors.forename: Koichi
authors.initials: K
authors.lastname: Abe
authors.forename: Wataru
authors.initials: W
authors.lastname: Yoshida
authors.forename: Kotaro
authors.initials: K
authors.lastname: Terada
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yagi-Ishii
authors.forename: Stefano
authors.initials: S
authors.lastname: Ferri
authors.forename: Kazunori
authors.initials: K
authors.lastname: Ikebukuro
authors.forename: Koji
authors.initials: K
authors.lastname: Sode
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D005948:Glucose Dehydrogenases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D016297:Mutagenesis, Site-Directed; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity
chemical_list: D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone)