Biochemical studies of the actions of ethanol on acetylcholinesterase activity: ethanol-enzyme-solvent interaction.
1. Biochemical studies of the actions of ethanol on the activity of acetylcholinesterase (AChE), isolated from electric eel (Electrophorus electricus) and purified by affinity chromatography, were performed to elucidate ethanol-enzyme-solvent interactions. 2. Ethanol at a low concentration [( EtOH] = 2.7-200 mM) was found to enhance AChE activity slightly and systematically. 3. This observation was consistent with the result from enzyme-kinetic studies that ethanol might noncompetitively activate AChE activity at this lower concentration range. 4. If ethanol alters the hydrophobic site interaction on the enzyme and subsequently induces a favorable conformation for the active center of the enzyme, then a slight increase in the AChE activity in the presence of a low concentration of ethanol will be observed. 5. This speculation was supported by the finding of ethanol's ability to perturb the inhibition of AChE activity by tetrabutylammonium bromide and to affect hydrophobic interaction between this salt and AChE, as investigated by enzyme activity and microcalorimetric measurements. 6. The ethanol effect on the activity of this soluble AChE was found to be distinguishable from that on a membrane-bound AChE. 7. Furthermore, to elucidate the effect of ethanol-solvent interaction on AChE activity, enzyme activity in the presence of much higher concentrations of ethanol was also examined. 8. At [EtOH] greater than 800 mM, ethanol can perturb the structure of water around hydrophobic areas of AChE, causing an instability in the enzyme conformation and subsequently decreasing AChE activity.
Shin
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Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany 12201-0509.
Wu
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Chen
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D000110:Acetylcholinesterase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D002846:Chromatography, Affinity; D004593:Electrophorus; D004789:Enzyme Activation / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D007700:Kinetics; D011487:Protein Conformation; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D012995:Solubility; D012997:Solvents
D002800:Cholinesterase Inhibitors; D000644:Quaternary Ammonium Compounds; D012997:Solvents; D000431:Ethanol; C009405:tetrabutylammonium; D000110:Acetylcholinesterase