Aging Pathways for Organophosphate-Inhibited Human Butyrylcholinesterase, Including Novel Pathways for Isomalathion, Resolved by Mass Spectrometry.

He Li; Lawrence M. Schopfer; Florian Nachon; Marie-Thérèse Froment; Patrick Masson; Oksana Lockridge

Some organophosphorus compounds are toxic because they inhibit acetylcholinesterase (AChE) by phosphylation of the active site serine, forming a stable conjugate: Ser–O–P(O)–(Y)–(XR) (where X can be O, N, or S and Y can be methyl, OR, or SR). The inhibited enzyme can undergo an aging process, during which the X–R moiety is dealkylated by breaking either the P–X or the X–R bond depending on the specific compound, leading to a nonreactivatable enzyme. Aging mechanisms have been studied primarily using AChE. However, some recent studies have indicated that organophosphate-inhibited butyrylcholinesterase (BChE) may age through an alternative pathway. Our work utilized matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry to study the aging mechanism of human BChE inhibited by dichlorvos, echothiophate, diisopropylfluorophosphate (DFP), isomalathion, soman, sarin, cyclohexyl sarin, VX, and VR. Inhibited BChE was aged in the presence of H2O18 to allow incorporation of 18O, if cleavage was at the P–X bond. Tryptic-peptide organophosphate conjugates were identified through peptide mass mapping. Our results showed no aging of VX- and VR-treated BChE at 25°C, pH 7.0. However, BChE inhibited by dichlorvos, echothiophate, DFP, soman, sarin, and cyclohexyl sarin aged exclusively through O–C bond cleavage, i.e., the classical X–R scission pathway. In contrast, isomalathion aged through both X–R and P–X pathways; the main aged product resulted from P–S bond cleavage and a minor product resulted from O–C and/or S–C bond cleavage.

ORGANOPHOSPHORUS compounds; TOXICOLOGICAL interactions; ACETYLCHOLINESTERASE; AGE factors in pharmacokinetics

Toxicological Sciences, 2007, Vol 100, Issue 1, p136

1096-6080

Academic Journal

10.1093/toxsci/kfm215