Chemical modification of acetylcholinesterase from eel and basal ganglia: effect on the acetylcholinesterase and aryl acylamidase activities.
The effect of chemical modification on the acetylcholinesterase and the aryl acylamidase activities of purified acetylcholinesterase from electric eel and basal ganglia was investigated in the presence and absence of acetylcholine, the substrate of acetylcholinesterase, and 1,5-bis[4-(allyldimethylammonium)phenyl]pentan-3-one dibromide (BW284C51), a reversible competitive inhibitor of acetylcholinesterase. Trinitrobenzenesulfonic acid, pyridoxal phosphate, acetic anhydride, diethyl pyrocarbonate, and 2-hydroxy-5-nitrobenzyl bromide under specified conditions inactivated both acetylcholinesterase and aryl acylamidase in the absence of acetylcholine and BW284C51. Chemical modifications in the presence of acetylcholine and BW284C51 by all the above except diethyl pyrocarbonate selectively prevented the loss of acetylcholinesterase but not aryl acylamidase activity; modification by diethyl pyrocarbonate in the presence of acetylcholine and BW284C51 prevented the loss of both acetylcholinesterase and aryl acylamidase activities. Treatment with N-acetylimidazole resulted in the inactivation of acetylcholinesterase and the activation of aryl acylamidase. These changes in both the activities could be prevented by acetylcholine and BW284C51. Modification by phenylglyoxal, 2,4-pentanedione, or N-ethylmaleimide did not affect the enzyme activities. Indophenylacetate hydrolase activity followed a pattern similar to that of acetylcholinesterase in all the above modification studies. The results suggested essential lysine, tyrosine, tryptophan, and histidine residues for the active center of acetylcholinesterase and essential lysine, histidine, and tryptophan residues for the active center of aryl acylamidase.(ABSTRACT TRUNCATED AT 250 WORDS)
D000110:Acetylcholinesterase / Q000378:metabolism*; D000581:Amidohydrolases / Q000378:metabolism*; D000818:Animals; D000936:Antigen-Antibody Complex; D001479:Basal Ganglia / Q000201:enzymology*; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D004593:Electrophorus; D010450:Endopeptidases; D007093:Imidazoles / Q000494:pharmacology; D007106:Immune Sera; D007700:Kinetics; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D012701:Serotonin / Q000494:pharmacology; D012756:Sheep; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology
D000936:Antigen-Antibody Complex; D007093:Imidazoles; D007106:Immune Sera; D013439:Sulfhydryl Reagents; D012701:Serotonin; D011732:Pyridoxal Phosphate; D014302:Trinitrobenzenesulfonic Acid; D000110:Acetylcholinesterase; D010450:Endopeptidases; D000581:Amidohydrolases; C019732:aryl acylamidase; D004047:Diethyl Pyrocarbonate; C012160:N-acetylimidazole