ORGANIC
LETTERS
2011
Vol. 13, No. 11
2799–2801
Ratiometric Fluorescence Imaging
of Cellular Glutathione
Gun-Joong Kim,† Kiwon Lee,‡ Hyockman Kwon,‡ and Hae-Jo Kim*,†
Department of Chemistry and Department of Bioscience and Biotechnology, Hankuk
University of Foreign Studies, Yongin 449-791, Republic of Korea
Received February 17, 2011
ABSTRACT
A fluorescent probe (1) with a hydrogen bond was designed for the detection of GSH. The probe exhibited a rapid and ratiometric fluorescence
response to GSH through a Michael reaction and allowed us to obtain clear cellular images for GSH.
Biothiols such as cysteine (Cys), homocysteine (Hcy),
and glutathione (GSH) are involvedinmyriadvital cellular
processes including redox homeostasis1 and cellular
growth.2 Alteration of the cellular biothiols is also impli-
cated in cancer and AIDS.3 Although a number of cell-
permeable, synthetic probes have been developed on the
basis of the changes in fluorescence intensity upon binding
with the biothiols,4 they cannot provide quantitative
information about changes in biothiol concentrations
caused by variations in excitation intensity, emission col-
lection efficiency, and artifacts associated with probe con-
centration and the environment. Therefore, it is of great
scientific interest and intense activity to develop a ratio-
metric probe that exhibits a fast5 and large shift6 in
emission or excitation profiles upon complexation with
biothiols. Herein we report a designed fluorescent probe
(1) for biothiols that is highly activated by an intramole-
cular H-bond and exhibits a rapid and ratiometric re-
sponse both in vitro and in vivo.
† Department of Chemistry.
‡ Department of Bioscience and Biotechnology.
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Recently, we reported optical probes with an o-hydro-
xy carbonyl group for effective intramolecular hydrogen
bonding that could be applied for the detection of such a
reactive nucleophile as cyanide or amino acid including
Hcy.7 In this line of research, we prepared a coumarin
derivative (1) possessing a hydrogen-bonded enone unit
(Scheme 1) where the coumarin moiety was designed to
function as a signaling unit, the conjugated enone as a
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J.-I. Chem. Commun. 2008, 6173.
r
10.1021/ol200967w
2011 American Chemical Society
Published on Web 05/06/2011