Abstract
Acetylcholinesterase (AChE) (EC 3.1.1.7) was modified with activated monomethoxypolyethylene glycol (mPEG). A decrease of 50% in the catalytic activity was measured during the coupling reaction and the change in the surface properties of AChE was used to separate by hydrophobic interaction chromatography the native and the modified enzyme. The native and the modified enzymes were found to have the same optimalcatalytic conditions. Moreover, the Michaelis constant of both enzymes were similar, whereas theV m and the bimolecular-velocity constant calculated for organophosphorus inhibitors were slightly higher for the modified AChE. Finally, the modification with mPEG did not improve the thermal stability, whereas the stability in a few organic solvents increased.
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Abbreviations
- AChE:
-
acetylcholinesterase
- ASChl:
-
acetylthiocholineiodide
- BCA:
-
Bicinchoninic acid
- DTNB:
-
5,5′-dithiobis (2-nitrobenzoÏcacid)
- ki:
-
bimolecular velocity constant
- Kmapp:
-
Apparent Michaelis constant
- mPEG:
-
methoxypolyethylene glycol
- mPEGpn:
-
methoxypolyethylene glycol pnitrophenyl carbonate
- PE:
-
parathion ethyl
- Px:
-
paraoxon ethyl
- Vm:
-
maximal velocity
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Garcia, D., Marty, JL. Chemical modification of acetylcholinesterase with methoxypolyethylene glycol. Appl Biochem Biotechnol 67, 153–163 (1997). https://doi.org/10.1007/BF02787849
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DOI: https://doi.org/10.1007/BF02787849