Design and synthesis of an enzyme activity-based labeling molecule with fluorescence spectral change.
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.
Komatsu
Toru
T
Graduate School of Pharmaceutical Sciences, the University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Kikuchi
Kazuya
K
Takakusa
Hideo
H
Hanaoka
Kenjiro
K
Ueno
Tasuku
T
Kamiya
Mako
M
Urano
Yasuteru
Y
Nagano
Tetsuo
T
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*
D005456:Fluorescent Dyes; D001616:beta-Galactosidase