title: Design and synthesis of an enzyme activity-based labeling molecule with fluorescence spectral change.
abstract: Methods of covalent labeling of a specific tag protein with small-molecular dyes play an important role in studying dynamic behaviors of proteins in living cells. On the basis of quinone methide chemistry, we designed and synthesized a beta-galactosidase labeling probe, CMFbeta-gal, which shows a fluorescence wavelength change accompanying the labeling reaction, owing to fluorescence resonance energy transfer (FRET). Since the FRET efficiency changes accompanying the labeling reaction, fluorescence of labeled protein can be observed separately from that of the unreacted probe, so immediate detection of the target protein is possible. This is the first report of a protein labeling probe which features a change of fluorescence wavelength upon reaction, allowing the labeled protein to be detected even in the presence of unreacted probe.
authors.affiliation: Graduate School of Pharmaceutical Sciences, the University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Komatsu
authors.forename: Kazuya
authors.initials: K
authors.lastname: Kikuchi
authors.forename: Hideo
authors.initials: H
authors.lastname: Takakusa
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Hanaoka
authors.forename: Tasuku
authors.initials: T
authors.lastname: Ueno
authors.forename: Mako
authors.initials: M
authors.lastname: Kamiya
authors.forename: Yasuteru
authors.initials: Y
authors.lastname: Urano
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nagano
mesh_terms: D002460:Cell Line; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry*; D006801:Humans; D015394:Molecular Structure; D013050:Spectrometry, Fluorescence; D001616:beta-Galactosidase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D001616:beta-Galactosidase