Evolution of fluorescein as a platform for finely tunable fluorescence probes.
Fluorescence imaging is the most powerful technique currently available for continuous observation of dynamic intracellular processes in living cells. Suitable fluorescence probes are naturally of critical importance for fluorescence imaging, but only a very limited range of biomolecules can currently be visualized because of the lack of flexible design strategies for fluorescence probes. At present, design is largely empirical. Here we show that the carboxylic group of traditional fluorescein dyes, formerly considered indispensable, has been replaced with other substituents, affording various kinds of new fluoresceins. Further, by breaking out of the traditional structure of fluorescein, we developed the first and totally rational design strategy for novel fluorescence probes based on a strict photochemical basis. The value of this approach is exemplified by its application to develop a novel, highly sensitive, and membrane-permeable fluorescence probe for beta-galactosidase, which is the most widely used reporter enzyme.
Urano
Yasuteru
Y
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. urano@mol.f.u-tokyo.ac.jp
Kamiya
Mako
M
Kanda
Kojiro
K
Ueno
Tasuku
T
Hirose
Kenzo
K
Nagano
Tetsuo
T
D015374:Biosensing Techniques / Q000379:methods; D005452:Fluoresceins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D011789:Quantum Theory; D013329:Structure-Activity Relationship; D001616:beta-Galactosidase / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics
D005452:Fluoresceins; D005456:Fluorescent Dyes; D001616:beta-Galactosidase