A novel fluorogenic probe for the investigation of free thiols: Application to kinetic measurements of acetylcholinesterase activity

[Display omitted] •A novel coumarin-derived turn-on probe for the detection of thiols with sufficient solubility in aqueous environments and excellent fluorescence characteristics.•A fluorescence-based assay for acetylcholinesterase activity as an addition to the established Ellman methodology.•Eval...

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Bibliographic Details
Published in:Toxicology letters 2016-02, Vol.244, p.161-166
Main Authors: Mertens, Matthias D., Bierwisch, Anne, Li, Tianwei, Gütschow, Michael, Thiermann, Horst, Wille, Timo, Elsinghorst, Paul W.
Format: Article
Language:English
Subjects:
Acetylcholinesterase
Acetylcholinesterase - metabolism
Activity assay
Algorithms
Aqueous environments
Asymmetry
Cleavage
Coumarins - metabolism
Fluorescence
Fluorescent Dyes - metabolism
GPI-Linked Proteins - metabolism
Humans
Kinetics
Least-Squares Analysis
Mathematical analysis
Nonlinear Dynamics
Quenching
Signal Processing, Computer-Assisted
Solubility
Spectrometry, Fluorescence
Sulfhydryl Compounds - metabolism
Thiols
Turn-on probe
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Summary:[Display omitted] •A novel coumarin-derived turn-on probe for the detection of thiols with sufficient solubility in aqueous environments and excellent fluorescence characteristics.•A fluorescence-based assay for acetylcholinesterase activity as an addition to the established Ellman methodology.•Evaluation of the asymmetric sigmoidal product formation curves applying a three parameter Gompertz equation. A novel coumarin-derived thiol probe, based on the thiol-promoted cleavage of a quenching 2,4-dinitrobenzenesulfonyl group is described. The probe shows a sensitive fluorescence turn-on and sufficient solubility in aqueous environments. As a proof of concept, a new assay for AChE activity was developed as a useful addition to the established Ellman method. The observed reaction kinetics followed an asymmetric sigmoidal pattern and were successfully evaluated applying a three parameter Gompertz equation. Providing a linear relationship between the detected fluorescence formation curves and corresponding enzyme activities, this probe appears as a valuable tool for AChE activity measurements.
ISSN:0378-4274
1879-3169
DOI:10.1016/j.toxlet.2015.10.012