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Cited by 254

The DOI matches its Crossref record; 59 of the 59 references it cites were found in a registry or an index.

Highly Selective Off–On Fluorescent Probe for Imaging Thioredoxin Reductase in Living Cells

Open access
Zhang, Liangwei, Duan, Dongzhu, Liu, Yaping, Ge, Chunpo, Cui, Xuemei, Sun, Jinyu, Fang, Jianguo·2014·Journal of the American Chemical Society
DOI 10.1021/ja408792k
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Abstract

The first fluorescent probe for mammalian thioredoxin reductase (TrxR), TRFS-green, was designed, synthesized, and fully evaluated. The probe features a 1,2-dithiolane scaffold with a quenched naphthalimide fluorophore. TRFS-green displays a green fluorescence off-on change induced by the TrxR-mediated disulfide cleavage and subsequent intramolecular cyclization to liberate the masked naphthalimide fluorophore. It was demonstrated in vitro that TRFS-green manifests high selectivity toward TrxR over other related enzymes and various small molecule thiols as well as biological reducing molecules. HPLC analyses indicated that TRFS-green was exclusively converted to naphthalimide catalyzed by TrxR. The ability in triggering on the fluorescence signal by cellular protein extracts correlates well with the endogenous TrxR activity in different cells. Furthermore, inhibition of TrxR by 2,4-dinitrochlorobenzene or depletion of TrxR by immunoprecipitation remarkably decreases the reduction of TRFS-green by cellular protein extracts. Finally, TRFS-green was successfully applied in imaging TrxR activity in living cells. The fluorescence signal of TRFS-green in living cells was inhibited by pretreating the cells with TrxR inhibitor in a dose-dependent manner, potentiating the development of living cell-based screening assay for identifying TrxR inhibitors. We expect the novel fluorescent probe TRFS-green would facilitate the discovery of TrxR-targeting small molecules for potential therapeutic agents and provide significant advances in understanding the physiological/pathophysiological functions of TrxR in vivo.

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Metadata

Authors
Zhang, L., Duan, D., Liu, Y., Ge, C., Cui, X., Sun, J., & Fang, J.
Venue
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Published
2013-12-18
Volume
136
Issue
1
Pages
226-233
ISSN
  • 0002-7863 (ISSN-L)
  • 1520-5126
DOI
10.1021/ja408792k
OpenAlex
W2017648151
Cited by
254
References
59
FWCI
6.46
Type
journal article
Language
en
Open access
green

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