期刊
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
卷 144, 期 1, 页码 174-183出版社
AMER CHEMICAL SOC
DOI: 10.1021/jacs.1c07954
关键词
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资金
- China Scholarship Council
- University of Bath
- EPSRC Centre for Doctoral Training in Catalysis [EP/L016443/1]
- EPSRC
- Royal Society
- Open Research Fund of the School of Chemistry and Chemical Engineering, Henan Normal University [2020ZD01]
- National Natural Science Foundation of China [21927811, 22074083, 21672150, 21302125, 21722501, 11974103]
- Key Research and Development Program of Shandong Province [2018YFJH0502]
- National Science Foundation of Shandong Province of China [ZR2020ZD17]
- Alexander von Humboldt Foundation (AvH), Shanghai Rising-Star Program [19QA1406400]
- Shanghai Government [18DZ2254200]
- China Postdoctoral Science Foundation [2020M681196]
- National Natural Science Foundation [91853201]
- Shanghai Municipal Science and Technology Major Project [2018SHZDZX03]
- National Key Sci-Tech Special Projects of Infection Diseases of China [2018ZX10732202]
- International Cooperation Program of Shanghai Science and Technology Committee [17520750100]
- Zhongyuan High Level Talents Special Support Plan-Science and Technology Innovation Leading Talents [204200510006]
- Program for Science Technology Innovation Talents in Universities of Henan Province [21HASTIT019]
- Robert A. Welch Foundation [F-0018]
The fluorescent probe ATP-LW was developed for simultaneous detection of ONOO- and ATP levels. By utilizing different emission properties of the products, ONOO- levels can be monitored in the green channel and ATP concentrations in the red channel. The probe was successfully used in hepatocytes to demonstrate changes in signal intensity in response to ATP synthase inhibition and exogenous ONOO- exposure.
Changes in adenosine triphosphate (ATP) and peroxynitrite (ONOO-) concentrations have been correlated in a number of diseases including ischemia-reperfusion injury and drug-induced liver injury. Herein, we report the development of a fluorescent probe ATP-LW, which enables the simultaneous detection of ONOO- and ATP. ONOO- selectively oxidizes the boronate pinacol ester of ATP-LW to afford the fluorescent 4-hydroxy-1,8-naphthalimide product NA-OH (lambda(ex) = 450 nm, lambda(em) = 562 nm or lambda(ex) = 488 lambda(em) = 568 nm). In contrast, the binding of ATP to ATP-LW induces the spirolactam ring opening of rhodamine to afford a highly emissive product (lambda(ex )= 520 nm, lambda(em) = 587 nm). Due to the differences in emission between the ONOO- and ATP products, ATP-LW allows ONOO- levels to be monitored in the green channel (lambda(ex) = 488 nm, lambda(em) = 500-575 nm) and ATP concentrations in the red channel (lambda(ex) = 514 nm, lambda(em) = 575-650 nm). The use of ATP-LW as a combined ONOO- and ATP probe was demonstrated using hepatocytes (HL-7702 cells) in cellular imaging experiments. Treatment of HL-7702 cells with oligomycin A (an inhibitor of ATP synthase) resulted in a reduction of signal intensity in the red channel and an increase in that of the green channel as expected for a reduction in ATP concentrations. Similar fluorescence changes were seen in the presence of SIN-1 (an exogenous ONOO- donor).
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