Screening of acetylcholinesterase inhibitors in snake venom by electrospray mass spectrometry

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Liesener, André and Perchuc, Anna-Maria and Schöni, Reto and Schebb, Nils Helge and Wilmer, Marianne and Karst, Uwe (2007) Screening of acetylcholinesterase inhibitors in snake venom by electrospray mass spectrometry. Pure and Applied Chemistry, 79 (12). pp. 2339-2349. ISSN 0033-4545

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Abstract:An electrospray ionization/mass spectrometry (ESI/MS)-based assay for the determination of acetylcholinesterase (AChE)-inhibiting activity in snake venom was developed. It allows the direct monitoring of the natural AChE substrate acetylcholine (AC) and the respective product choline. The assay scheme was employed in the screening for neurotoxic activity in fractions of the venom of Bothrops moojeni. AChE inhibition was assessed in two fractions. As a positive control, the established AChE inhibitor 1,5-bis(4-allyl-dimethylammoniumphenyl)pentan-3-one dibromide (BW284c51) was used, a dose-response curve for this compound was generated and the IC50 value for the inhibitor was determined to be 1.60 ± 0.09 × 10-9 mol L-1. The dose-response curve was used as "calibration function" for the venom inhibition activity, resulting in BW284c51-equivalent concentrations of 1.76 × 10-9 mol L-1 and 1.07 × 10-9 mol L-1 for the two fractions containing activity. The ESI/MS-based assay scheme was validated using the established Ellman reaction. The data obtained using both methods were found to be in good agreement. The ESI/MS-based assay scheme is therefore an attractive alternative to the standard colorimetric assay.
*Invited contribution to a collection of papers for the IUPAC project 2005-042-1-300 "Chemistry for Biology".
Item Type:Article
Copyright:© 2010 International Union of Pure and Applied Chemistry
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Link to this item:http://purl.utwente.nl/publications/74895
Official URL:http://dx.doi.org/10.1351/pac200779122339
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