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

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)136-145
Number of pages10
JournalToxicological Sciences
Volume100
Issue number1
DOIs
StatePublished - Nov 1 2007

Fingerprint

Butyrylcholinesterase
Sarin
Organophosphates
Mass spectrometry
Mass Spectrometry
Aging of materials
Dichlorvos
Soman
Isoflurophate
Acetylcholinesterase
Organophosphorus Compounds
Peptides
Peptide Mapping
Poisons
Enzymes
Serine
Ionization
Desorption
Catalytic Domain
Lasers

Keywords

  • Aging
  • Butyrylcholinesterase
  • Mass spectrometry
  • Organophosphate

ASJC Scopus subject areas

  • Toxicology

Cite this

Aging pathways for organophosphate-inhibited human butyrylcholinesterase, including novel pathways for isomalathion, resolved by mass spectrometry. / Li, He; Schopfer, Lawrence M; Nachon, Florian; Froment, Marie Thérèse; Masson, Patrick; Lockridge, Oksana.

In: Toxicological Sciences, Vol. 100, No. 1, 01.11.2007, p. 136-145.

Research output: Contribution to journalArticle

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