title: The development of new molecular tools containing a chemically synthesized carbohydrate ligand for the elucidation of carbohydrate roles via photoaffinity labeling: carbohydrate-protein interactions are affected by the structures of the glycosidic bonds and the reducing-end sugar.
abstract: Photoaffinity labeling technology is a highly efficient method for cloning carbohydrate-binding proteins. When the carbohydrate probes are synthesized according to conventional methods, however, the reducing terminus of the sugar is opened to provide an acyclic structure. Our continued efforts to solve this problem led to the development of new molecular tools with an oligosaccharide structure that contains a phenyldiazirine group for the elucidation of carbohydrate-protein interactions. We investigated whether carbohydrate-lectin interactions are affected by differences in the glycosidic formation and synthesized three types of molecular tools containing Galp-GlcpNAc disaccharide ligands and a photoreactive group (1, 2, 3). Photoaffinity labeling validated the recognition of the new ligand by different glycosidic bonds. Photoaffinity labeling also demonstrated that both the reducing end sugar and non-reducing end sugar recognized the Erythrina cristagalli agglutinin.
authors.affiliation: School of Pharmaceutical Sciences, Kyushu University of Health and Welfare, 1714-1 Yoshino-cho, Nobeoka City, Miyazaki 882-8508, Japan. Electronic address: ohtsuka@phoenix.ac.jp.
authors.forename: Isao
authors.initials: I
authors.lastname: Ohtsuka
authors.affiliation: Suzuka University of Medical Science, 3500-3 Minamitamagaki-cho, Suzuka City, Mie 513-8670, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Sadakane
authors.affiliation: Faculty of Pharmacy, Keio University, 1-5-30 Shibakoen, Minato-ku, Tokyo 105-8512, Japan.
authors.forename: Noriyasu
authors.initials: N
authors.lastname: Hada
authors.affiliation: School of Pharmaceutical Sciences, Kyushu University of Health and Welfare, 1714-1 Yoshino-cho, Nobeoka City, Miyazaki 882-8508, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Higuchi
authors.affiliation: School of Pharmaceutical Sciences, Kyushu University of Health and Welfare, 1714-1 Yoshino-cho, Nobeoka City, Miyazaki 882-8508, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Atsumi
authors.affiliation: School of Pharmaceutical Sciences, Kyushu University of Health and Welfare, 1714-1 Yoshino-cho, Nobeoka City, Miyazaki 882-8508, Japan.
authors.forename: Nobuko
authors.initials: N
authors.lastname: Kakiuchi
mesh_terms: D000373:Agglutinins / Q000737:chemistry / Q000378:metabolism; D002241:Carbohydrates / Q000138:chemical synthesis / Q000737:chemistry*; D004187:Disaccharides / Q000737:chemistry / Q000378:metabolism; D004895:Erythrina / Q000378:metabolism; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D019920:Photoaffinity Labels / Q000737:chemistry*; D011485:Protein Binding; D014466:Ultraviolet Rays
chemical_list: D000373:Agglutinins; D002241:Carbohydrates; D004187:Disaccharides; D037102:Lectins; D008024:Ligands; D019920:Photoaffinity Labels
keywords: Carbohydrate–lectin interactions; Different glycosidic bonds; Molecular tool; Oligosaccharide synthesis; Photoaffinity labeling