Polyproline promotes tetramerization of recombinant human butyrylcholinesterase

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23 Scopus citations


Human BChE (butyrylcholinesterase) protects against the toxicity of organophosphorus nerve agents and pesticides. BChE purified from human plasma is limited and pathogen carry-over is a concern. Unlike the native BChE tetrameric complex with a residence time of days, rBChE (recombinant BChE) is produced predominantly as dimers andmonomers that are cleared from the circulation within minutes. Assembly into tetramers requires incorporation of proline-rich peptides, a process that was thought to occur intracellularly. Our goal was to determine whether polyproline added to rBChE under cell-free conditions would promote tetramerization. Secreted rBChE was purified by procainamide affinity chromatography, and synthetic polyprolines (8-mer to 300-mer) were tested to determine their effect on tetramer assembly. These studies demonstrated that 90-98% of purified rBChE (65 μM) could be assembled into tetramers when incubated with synthetic 17-mer or 50-mer polyproline peptides (100 μM) for 1.5 h at 25°C. However, rBChE tetramerizationwas inefficient with smaller 8-mer polyproline peptides and larger 300-mer polyproline proteins. Collectively, these studies demonstrated that the eukaryotic cellularmachinery is not required for assembly of active BChE into tetramers and that this process can occur in vitro with purified rBChE in the presence of peptides containing 15-50 consecutive proline residues.

Original languageEnglish (US)
Pages (from-to)329-335
Number of pages7
JournalBiochemical Journal
Issue number2
StatePublished - Sep 1 2014


  • Enzyme stability
  • Human butyrylcholinesterase
  • Polyproline peptide
  • Recombinant expression
  • Tetrameric bioscavenger

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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