title: Complementary DNA display selection of high-affinity peptides binding the vacuolating toxin (VacA) of Helicobacter pylori.
abstract: Artificial peptides designed for molecular recognition of a bacterial toxin have been developed. Vacuolating cytotoxin A protein (VacA) is a major virulence factor of Helicobacter pylori, a gram-negative microaerophilic bacterium inhabiting the upper gastrointestinal tract, particularly the stomach. This study attempted to identify specific peptide sequences with high affinity for VacA using systematic directed evolution in vitro, a cDNA display method. A surface plasmon resonance-based biosensor and fluorescence correlation spectroscopy to examine binding of peptides with VacA identified a peptide (GRVNQRL) with high affinity. Cyclization of the peptide by attaching cysteine residues to both termini improved its binding affinity to VacA, with a dissociation constant (Kd ) of 58 nm. This study describes a new strategy for the development of artificial functional peptides, which are promising materials in biochemical analyses and medical applications.
authors.affiliation: Graduate School of Bioscience, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama, Shiga, 526-0829, Japan.
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Hayakawa
authors.affiliation: Graduate School of Bioscience, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama, Shiga, 526-0829, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Matsuno
authors.affiliation: Graduate School of Bioscience, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama, Shiga, 526-0829, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Tanaka
authors.affiliation: Institute for Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, Nagasaki, 852-8523, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Wada
authors.affiliation: JANUSYS Co., Ltd., Kamiaoki 3-12-18-508, Kawaguchi, Saitama, 333-0844, Japan.
authors.forename: Koichiro
authors.initials: K
authors.lastname: Kitamura
authors.affiliation: Lifetech Co., Ltd., Miyadera 4074, Iruma, Saitama, 358-0014, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Takei
authors.affiliation: Graduate School of Bioscience, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama, Shiga, 526-0829, Japan.
authors.forename: Ryuzo
authors.initials: R
authors.lastname: Sasaki
authors.affiliation: Graduate School of Bioscience, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama, Shiga, 526-0829, Japan.
authors.forename: Tamio
authors.initials: T
authors.lastname: Mizukami
authors.affiliation: Graduate School of Bioscience, Nagahama Institute of Bio-Science and Technology, Tamura 1266, Nagahama, Shiga, 526-0829, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Hasegawa
mesh_terms: D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics*; D016480:Helicobacter pylori / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D001427:Bacterial Toxins; D018076:DNA, Complementary; D010455:Peptides
keywords: Helicobacter pylori; artificial functional peptide; cDNA display method; in vitro peptide evolution; vacuolating cytotoxin A