title: Peptidoglycan microarray as a novel tool to explore protein-ligand recognition.
abstract: Peptidoglycan is a giant bag-shaped molecule essential for bacterial cell shape and resistance to osmotic stresses. The activity of a large number of bacterial surface proteins involved in cell growth and division requires binding to this macromolecule. Recognition of peptidoglycan by immune effectors is also crucial for the establishment of the immune response against pathogens. The availability of pure and chemically defined peptidoglycan fragments is a major technical bottleneck that has precluded systematic studies of the mechanisms underpinning protein-mediated peptidoglycan recognition. Here, we report a microarray strategy suitable to carry out comprehensive studies to characterize proteins-peptidoglycan interactions. We describe a method to introduce a functional group on peptidoglycan fragments allowing their stable immobilization on amorphous carbon chip plates to minimize nonspecific binding. Such peptidoglycan microarrays were used with a model peptidoglycan binding protein-the human peptidoglycan recognition protein-S (hPGRP-S). We propose that this strategy could be implemented to carry out high-throughput analyses to study peptidoglycan binding proteins. © 2016 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 106: 422-429, 2016.
authors.affiliation: Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka, 560-0043, Japan.
authors.forename: Ning
authors.initials: N
authors.lastname: Wang
authors.affiliation: HiPep Laboratories, 486-46 Nakatsukasa-Cho, Kamigyo-Ku, Kyoto, 602-8158, Japan.
authors.forename: Akiyoshi
authors.initials: A
authors.lastname: Hirata
authors.affiliation: HiPep Laboratories, 486-46 Nakatsukasa-Cho, Kamigyo-Ku, Kyoto, 602-8158, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Nokihara
authors.affiliation: Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka, 560-0043, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Fukase
authors.affiliation: Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka, 560-0043, Japan.|Department of Chemistry, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-Ku, Yokohama, Kanagawa, 223-8522, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Fujimoto
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry*; D006801:Humans; D010457:Peptidoglycan / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D010457:Peptidoglycan; C118897:peptidoglycan recognition protein
keywords: microarray; peptidoglycan; recognition