Assessment of acetylcholinesterase activity using indoxylacetate and comparison with the standard Ellman's method
Jazyk angličtina Země Švýcarsko Médium print-electronic
Typ dokumentu srovnávací studie, časopisecké články, práce podpořená grantem
PubMed
21731462
PubMed Central
PMC3127138
DOI
10.3390/ijms12042631
PII: ijms12042631
Knihovny.cz E-zdroje
- Klíčová slova
- 5,5′-dithio-bis-2-nitrobenzoic acid, Alzheimer’s disease, acetylcholinesterase, enzyme activity, indoxylacetate, nerve agents, oxime reactivator,
- MeSH
- acetylcholinesterasa metabolismus MeSH
- cholinesterasové inhibitory chemie metabolismus MeSH
- enzymatické testy * MeSH
- indoly chemie metabolismus MeSH
- kinetika MeSH
- kyselina dithionitrobenzoová chemie metabolismus MeSH
- obidoxim chlorid chemie metabolismus MeSH
- oximy chemie metabolismus MeSH
- paraoxon chemie metabolismus MeSH
- pralidoximové sloučeniny chemie metabolismus MeSH
- pyridinové sloučeniny chemie metabolismus MeSH
- reaktivátory cholinesterázy chemie metabolismus MeSH
- substrátová specifita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- acetylcholinesterasa MeSH
- asoxime chloride MeSH Prohlížeč
- cholinesterasové inhibitory MeSH
- indoly MeSH
- indoxyl acetate MeSH Prohlížeč
- kyselina dithionitrobenzoová MeSH
- obidoxim chlorid MeSH
- oximy MeSH
- paraoxon MeSH
- pralidoxime MeSH Prohlížeč
- pralidoximové sloučeniny MeSH
- pyridinové sloučeniny MeSH
- reaktivátory cholinesterázy MeSH
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 V(max) (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.
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