title: Photochemical construction of coumarin fluorophore on affinity-anchored protein.
abstract: A simple and unique construction of a coumarin fluorophore on a prey protein has been achieved by sequential photoreactions using a nonfluorescent bait protein. The bait protein was first modified with a novel diazirine-based photo-cross-linker containing an o-hydroxycinnamoyl unit. The strategy involves two wavelength-dependent photoreactions: photolysis of the diazirine group and E to Z photoisomerization of the cinnamoyl group. The cross-linked interacting partners were cleaved by the consecutive steps of photoisomerization and thermal lactonization accompanied with the creation of a coumarin derivative on the prey protein as a fluorescent tag. Finally, the methodology was successfully applied for coumarin labeling of antilysozymes expressed on living B cells by using photoactivatable lysozymes.
authors.affiliation: Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930-0194, Japan. yasu@pha.u-toyama.ac.jp
authors.forename: Takenori
authors.initials: T
authors.lastname: Tomohiro
authors.forename: Kenichi
authors.initials: K
authors.lastname: Kato
authors.forename: Souta
authors.initials: S
authors.lastname: Masuda
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kishi
authors.forename: Yasumaru
authors.initials: Y
authors.lastname: Hatanaka
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D003374:Coumarins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D051379:Mice; D009113:Muramidase / Q000276:immunology; D055668:Photochemical Processes; D011506:Proteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D000906:Antibodies; D003374:Coumarins; D005456:Fluorescent Dyes; D011506:Proteins; C030123:coumarin; D009113:Muramidase