Assessment of Acetylcholinesterase Activity Using Indoxylacetate and Comparison with the Standard Ellman’s Method
Open Access
- 18 April 2011
- journal article
- research article
- Published by MDPI AG in International Journal of Molecular Sciences
- Vol. 12 (4), 2631-2640
- https://doi.org/10.3390/ijms12042631
Abstract
Assay of acetylcholinesterase (AChE) activity plays an important role in diagnostic, detection of pesticides and nerve agents, in vitro characterization of toxins and drugs including potential treatments for Alzheimer’s disease. These experiments were done in order to determine whether indoxylacetate could be an adequate chromogenic reactant for AChE assay evaluation. Moreover, the results were compared to the standard Ellman’s method. We calculated Michaelis constant Km (2.06 × 10−4 mol/L for acetylthiocholine and 3.21 × 10−3 mol/L for indoxylacetate) maximum reaction velocity Vmax (4.97 × 10−7 kat for acetylcholine and 7.71 × 10−8 kat for indoxylacetate) for electric eel AChE. In a second part, inhibition values were plotted for paraoxon, and reactivation efficacy was measured for some standard oxime reactivators: obidoxime, pralidoxime (2-PAM) and HI-6. Though indoxylacetate is split with lower turnover rate, this compound appears as a very attractive reactant since it does not show any chemical reactivity with oxime antidots and thiol used for the Ellman’s method. Thus it can be advantageously used for accurate measurement of AChE activity. Suitability of assay for butyrylcholinesterase activity assessment is also discussed.Keywords
This publication has 27 references indexed in Scilit:
- Galangin, a flavonol derived from Rhizoma Alpiniae Officinarum, inhibits acetylcholinesterase activity in vitroChemico-Biological Interactions, 2010
- In vitroreactivation of trichlorfon-inhibited butyrylcholinesterase using HI-6, obidoxime, pralidoxime and K048Journal of Enzyme Inhibition and Medicinal Chemistry, 2009
- Nerve Agents Assay Using Cholinesterase Based BiosensorElectroanalysis, 2009
- Stabilization of Torpedo californica Acetylcholinesterase by Reversible InhibitorsBiochemistry, 2008
- Diagnosis of Intoxication by the Organophosphate VX: Comparison Between an Electrochemical Sensor and Ellman´s Photometric MethodSensors, 2008
- Photometric microplate assay for estimation of the efficacy of paraoxon-inhibited acetylcholinesterase reactivationJournal of Enzyme Inhibition and Medicinal Chemistry, 2008
- Cholinesterase-based dipstick assay for the detection of organophosphate and carbamate pesticidesAnalytica Chimica Acta, 2007
- Cholinesterase pseudo-activity, oximolysis, esterolysis, thiocholine ester hydrolysis by oximes: What's in a name?Toxicology Letters, 2007
- Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling methodNature Protocols, 2006
- Role of butyrylcholinesterase in canine tracheal smooth muscle functionFEBS Letters, 1990