Characterization of butyrylcholinesterase from porcine milk

Ashima Saxena, Tatyana Belinskaya, Lawrence M Schopfer, Oksana Lockridge

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Human butyrylcholinesterase (HuBChE) is under development for use as a pretreatment antidote against nerve agent toxicity. Animals are used to evaluate the efficacy of HuBChE for protection against organophosphorus nerve agents. Pharmacokinetic studies of HuBChE in minipigs showed a mean residence time of 267 h, similar to the half-life of HuBChE in humans, suggesting a high degree of similarity between BChE from 2 sources. Our aim was to compare the biochemical properties of PoBChE purified from porcine milk to HuBChE purified from human plasma. PoBChE hydrolyzed acetylthiocholine slightly faster than butyrylthiocholine, but was sensitive to BChE-specific inhibitors. PoBChE was 50-fold less sensitive to inhibition by DFP than HuBChE and 5-fold slower to reactivate in the presence of 2-PAM. The amino acid sequence of PoBChE determined by liquid chromatography tandem mass spectrometry was 91% identical to HuBChE. Monoclonal antibodies 11D8, mAb2, and 3E8 (HAH 002) recognized both PoBChE and HuBChE. Assembly of 4 identical subunits into tetramers occurred by noncovalent interaction with polyproline-rich peptides in PoBChE as well as in HuBChE, though the set of polyproline-rich peptides in milk-derived PoBChE was different from the set in plasma-derived HuBChE tetramers. It was concluded that the esterase isolated from porcine milk is PoBChE.

Original languageEnglish (US)
Pages (from-to)38-49
Number of pages12
JournalArchives of Biochemistry and Biophysics
Volume652
DOIs
StatePublished - Aug 15 2018

Fingerprint

Butyrylcholinesterase
Milk
Swine
Butyrylthiocholine
Acetylthiocholine
Plasma (human)
Isoflurophate
Antidotes
Peptides
Pharmacokinetics
Miniature Swine
Liquid chromatography
Esterases
Human Milk
Tandem Mass Spectrometry
Mass spectrometry
Toxicity
Liquid Chromatography
Animals
Half-Life

Keywords

  • Acetylcholinesterase
  • Butyrylcholinesterase
  • Hupresin
  • Polyproline
  • Porcine
  • Tetramer

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Characterization of butyrylcholinesterase from porcine milk. / Saxena, Ashima; Belinskaya, Tatyana; Schopfer, Lawrence M; Lockridge, Oksana.

In: Archives of Biochemistry and Biophysics, Vol. 652, 15.08.2018, p. 38-49.

Research output: Contribution to journalArticle

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