title: Comparison of the reactivity of carbohydrate photoaffinity probes with different photoreactive groups.
abstract: A judicious choice of photoreactive group is critical in successful photoaffinity labeling studies of small molecule-protein interactions. A set of carbohydrate-based photoaffinity probes was prepared to compare the effects of three major photoreactive groups on the efficiency and selectivity of crosslinking a binding protein with low affinity. We showed that, despite the low crosslinking yield, the diazirine probe displayed the high ligand-dependent reactivity consistent with the ideal mechanism of photoaffinity labeling. Moreover, we demonstrated that, among the three photoreactive groups, only the diazirine probe achieved highly selective crosslinking of a low-affinity binding protein in cell lysate.
authors.affiliation: Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Naka-cho, 2-24-16, Koganei, Tokyo 184-8588 (Japan). sakuraik@cc.tuat.ac.jp.
authors.forename: Kaori
authors.initials: K
authors.lastname: Sakurai
authors.forename: Shimpei
authors.initials: S
authors.lastname: Ozawa
authors.forename: Rika
authors.initials: R
authors.lastname: Yamada
authors.forename: Tomoki
authors.initials: T
authors.lastname: Yasui
authors.forename: Sakae
authors.initials: S
authors.lastname: Mizuno
mesh_terms: D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism*; D003978:Diazomethane / Q000737:chemistry / Q000378:metabolism*; D019887:Peanut Agglutinin / Q000378:metabolism*; D019920:Photoaffinity Labels / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D014466:Ultraviolet Rays
chemical_list: D002241:Carbohydrates; D003432:Cross-Linking Reagents; D019887:Peanut Agglutinin; D019920:Photoaffinity Labels; D003978:Diazomethane
keywords: binding proteins; carbohydrates; chemical probes; photoaffinity labeling; photoreactive groups