A novel fluorogenic probe for the investigation of free thiols: Application to kinetic measurements of acetylcholinesterase activity. (26th February 2016)

• Record Type: Journal Article
• Authors: Mertens, Matthias D.; Bierwisch, Anne; Li, Tianwei; Gütschow, Michael; Thiermann, Horst; Wille, Timo; Elsinghorst, Paul W.
• Is Part Of: Toxicology letters. Volume 244(2016)
• Page Start: 161
• Page End: 166
• Publication Date: 2016-02-26
• Subjects: Thiols -- Turn-on probe -- Fluorescence -- Acetylcholinesterase -- Activity assay; Toxicology -- Periodicals; 363.179
• DOI: 10.1016/j.toxlet.2015.10.012
• Languages: English
• ISSNs: 0378-4274

Abstract

Graphical abstract: Highlights: 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. Abstract: 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.