Atypical effect of some spin trapping agents: reversible inhibition of acetylcholinesterase.
N-tert-butyl-alpha-phenylnitrone (PBN), a widely used nitrone-based free radical trap was recently shown to prevent acetylcholinesterase (AChE) inhibitors induced muscle fasciculations and brain seizures while being ineffective against glutamergic or cholinergic receptor agonist induced seizures. In the present study we compared the effects on AChE activity of four free radical spin traps PBN, alpha-(4-pyridil-1)-N-tert-butyl nitrone (POBN), N-tert-butyl-alpha-(2-sulfophenyl)-nitrone (S-PBN) and 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide (DEPMPO). The kinetics of AChE inhibition were studied in vitro using a spectrophotometric kinetic assay with AChE from rat brain, diaphragm, electric eel and mouse brain. Spin trapping compounds S-PBN and DEPMPO, in concentrations up to 3 mM did not inhibit hydrolysis of ACh, while PBN and POBN inhibited hydrolysis of ACh in a reversible and concentration-dependent manner. Double reciprocal plots of the reaction velocity against varying ACh concentrations at each inhibitor concentration were linear and generally indicated mixed type inhibition. PBN was the most potent inhibitor of mouse AChE with Ki and Ki' of 0.58 and 2.99 mM, respectively, and the weakest inhibitor of electric eel AChE. In contrast, POBN showed the highest affinity for electric eel enzyme, with Ki and Ki' values of 1.065 and 3.15 mM, respectively. These findings suggest that the effect of PBN and POBN on AChE activity does not depend on trapping of damaging reactive oxygen and that in addition to their antioxidant action other pharmacological effects of these compounds should be considered when neuroprotective actions of PBN or POBN are investigated.
Milatovic
D
D
Department of Pharmacology, Vanderbilt University, School of Medicine, Nashville, TN, USA.
Radic
Z
Z
Zivin
M
M
Dettbarn
W D
WD
D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D001557:Benzenesulfonates / Q000494:pharmacology*; D001921:Brain / Q000201:enzymology; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D003497:Cyclic N-Oxides / Q000494:pharmacology*; D003964:Diaphragm / Q000201:enzymology; D004593:Electrophorus; D016166:Free Radical Scavengers / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D018482:Muscle, Skeletal / Q000201:enzymology; D009589:Nitrogen Oxides / Q000494:pharmacology*; D011725:Pyridines; D051381:Rats; D017207:Rats, Sprague-Dawley; D013113:Spin Labels
D001557:Benzenesulfonates; D002800:Cholinesterase Inhibitors; D003497:Cyclic N-Oxides; D016166:Free Radical Scavengers; D009589:Nitrogen Oxides; D011725:Pyridines; D013113:Spin Labels; C043438:alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone; C094272:N-tert-butyl-(2-sulfophenyl)nitrone; C098505:5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide; C029217:phenyl-N-tert-butylnitrone; D000110:Acetylcholinesterase