A fluorescence-based assay for measuring the redox potential of 5-lipoxygenase inhibitors

The activities and side effects of 5-lipoxygenase (5-LO) inhibitors can be predicted by identifying their redox mechanisms. In this study, we developed a fluorescence-based method to measure the redox potential of 5-LO inhibitors and compared it to the conventional, absorbance-based method. After th...

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Bibliographic Details
Published in:PloS one 2014-02, Vol.9 (2), p.e87708
Main Authors: Lee, Sangchul, Park, Youngsam, Kim, Junghwan, Han, Sung-Jun
Format: Article
Language:English
Subjects:
Absorbance
Acids
Arachidonate 5-lipoxygenase
Arachidonate 5-Lipoxygenase - chemistry
Assaying
Asthma
Biological Assay - methods
Biology
Chemistry
Comparative analysis
Dose-Response Relationship, Drug
Electrode potentials
Enzymes
Fluoresceins - analysis
Fluorescence
Fluorescent Dyes - analysis
Humans
Inhibitors
Ligands
Lipid Peroxides - analysis
Lipids
Lipoxygenase
Lipoxygenase Inhibitors - analysis
Lipoxygenase Inhibitors - chemistry
Measurement
Medicine
Oxidation
Oxidation-Reduction
Peroxidase
Peroxide
Redox potential
Redox properties
Side effects
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Summary:The activities and side effects of 5-lipoxygenase (5-LO) inhibitors can be predicted by identifying their redox mechanisms. In this study, we developed a fluorescence-based method to measure the redox potential of 5-LO inhibitors and compared it to the conventional, absorbance-based method. After the pseudo-peroxidase reaction, the amount of remaining lipid peroxide was quantified using the H2DCFDA (2',7'-dichlorodihydrofluorescein diacetate) fluorescence dye. Our method showed large signal windows and provided comparable redox potential values. Importantly, the redox mechanisms of known inhibitors were accurately measured with the fluorescence assay, whereas the conventional, absorbance-based method showed contradictory results. Our findings suggest that our developed method is a better alternative for classifying the redox potential of 5-LO inhibitors, and the fluorescence assay can be effectively used to study the mechanisms of action that are related to redox cycling.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0087708