Smart Dual Quenching Strategy Enhances the Detection Sensitivity of Intracellular Furin

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Abstract
Development of sensitive fluorescence “Turn-On” strategies for imaging enzyme activity in living cells is of disease-diagnostic importance but remains challenging. Herein, by employing a click condensation reaction and rational design of a single quenched probe Cys(StBu)-Lys(Gly-Lys(DABCYL)-Gly-Gly-Arg-Arg-Val-Arg-Gly-FITC)-CBT (1), we developed a “smart” dual quenching strategy and applied it to detect intracellular furin activity with enhanced sensitivity. At physiological condition, 1 was subjected to reduction-controlled condensation reaction to form 1-NPs and its fluorescence intensity further dropped to 1/2.8 of its original. Upon furin cleavage in vitro, the dual quenched 1-NPs had fluorescence “Turn-On” contrast 11 folds more than that of single quenched control probe FITC-Gly-Arg-Val-Arg-Arg-Gly-Gly-Lys(DABCYL)-Gly-OH (1-P). Live cell imaging results indicated that 1 showed fluorescence “Turn-On” contrast 6.3-fold of that of 1-P for sensing intracellular furin activity. We envision that, by replacing the RRVR substrate with other enzyme-cleavable ones, our versatile “smart” dual quenching strategy could be easily adjusted for the detection (or imaging) of other intracellular enzymes’ activity with enhanced sensitivity.
Funding Information
  • Ministry of Science and Technology of the People's Republic of China (2016YFA0400904)
  • National Natural Science Foundation of China (21675145, 21725505)
  • Collaborative Innovation Center of Suzhou Nano Science and Technology
  • Hefei Center for Physical Science and Technology, Chinese Academy of Sciences (2016FXZY006)

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