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Self-Calibrating Bipartite Fluorescent Sensor for Nitroreductase Activity and Its Application to Cancer and Hypoxic
Shin A Yoon1, Jieun Chun2, Chulhun Kang2
1Department of Chemistry, Sookmyung Women's University, Seoul 04310, Korea.
ACS Applied Bio Materials
|January 11, 2022
Summary
Researchers developed a novel self-calibrating fluorescent probe to accurately measure nitroreductase (NTR) activity. This tool helps compare NTR levels in different cells, revealing higher activity in cancer cells and hypoxic states.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- Nitroreductases (NTRs) reduce aromatic nitro compounds to amino derivatives.
- Mammalian NTRs are largely uncharacterized but represent a therapeutic target for cancer and hypoxia.
- Fluorescent probes are used to assay NTR activity, but comparisons between cells are limited by nonenzymatic factors.
Purpose of the Study:
- To develop a self-calibrating fluorescent probe for accurate and comparable measurement of nitroreductase activity.
- To investigate the association of NTR activity with cancer cells and hypoxia.
Main Methods:
- Designed a bipartite fluorescent probe incorporating an NTR-sensitive nitronaphthalimide and a nonsensitive coumarin moiety.
- Utilized fluorescence ratio monitoring to quantify relative NTR activity.
- Applied the probe to compare NTR activity in noncancerous and cancerous cells.
Main Results:
- The bipartite probe enabled self-calibration, overcoming limitations of previous probes affected by nonenzymatic factors.
- Relative NTR activity could be reliably compared between different cell types.
- Demonstrated elevated NTR activity in cancerous cells and hypoxic conditions.
Conclusions:
- The developed self-calibrating probe is a valuable tool for accurate assessment of NTR activity.
- Elevated NTR activity is a potential biomarker for cancer and hypoxia.
- This probe facilitates comparative studies of NTR activity across various cellular contexts.
