Effect of different buffers on kinetic properties of human acetylcholinesterase and the interaction with organophosphates and oximes.
Acetylcholinesterase (AChE) is the primary target of organophosphorus compounds (OP). The investigation into interactions between AChE, OP and oximes in vitro may be affected by the experimental conditions, e.g. by the buffer system. Hence, it was tempting to investigate the Michaelis-Menten kinetics and the inhibition and reactivation kinetics of paraoxon-ethyl, sarin, soman and VX in the presence of phosphate, MOPS, Tyrode and TRIS buffer with human AChE. Compared to phosphate buffer, the inhibition and reactivation kinetics of human erythrocyte AChE were markedly changed by TRIS and in part by MOPS, whereas Tyrode showed similar results to phosphate buffer. These results indicate an effect of the tested buffers on the properties of AChE, and an interaction between OP and oximes has to be considered for the design of in vitro studies and may impair the comparison of data from different laboratories. In view of the comparability of human in vitro kinetic data determined with phosphate buffer with data from human OP poisoning, it seems to be a suitable buffer for the investigation into interactions between AChE, OP and oximes.
Wille
T
T
Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany.
Thiermann
H
H
Worek
F
F
D000110:Acetylcholinesterase / Q000187:drug effects / Q000378:metabolism* / Q000494:pharmacology; D002021:Buffers; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D004347:Drug Interactions; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D010755:Organophosphates / Q000494:pharmacology*; D010091:Oximes / Q000494:pharmacology*; D010261:Paraoxon / Q000031:analogs & derivatives / Q000494:pharmacology; D012524:Sarin / Q000494:pharmacology; D012999:Soman / Q000494:pharmacology
D002021:Buffers; D002800:Cholinesterase Inhibitors; D010755:Organophosphates; D010091:Oximes; C121104:ethylparaoxon; D012999:Soman; D012524:Sarin; D000110:Acetylcholinesterase; D010261:Paraoxon