Hydrolysis of oxo- and thio-esters by human butyrylcholinesterase

Patrick Masson, Marie Thérèse Froment, Emilie Gillon, Florian Nachon, Oksana Lockridge, Lawrence M Schopfer

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Catalytic parameters of human butyrylcholinesterase (BuChE) for hydrolysis of homologous pairs of oxo-esters and thio-esters were compared. Substrates were positively charged (benzoylcholine versus benzoylthiocholine) and neutral (phenylacetate versus phenylthioacetate). In addition to wild-type BuChE, enzymes containing mutations were used. Single mutants at positions: G117, a key residue in the oxyanion hole, and D70, the main component of the peripheral anionic site were tested. Double mutants containing G117H and mutations on residues of the oxyanion hole (G115, A199), or the π-cation binding site (W82), or residue E197 that is involved in stabilization of tetrahedral intermediates were also studied. A mathematical analysis was used to compare data for BuChE-catalyzed hydrolysis of various pairs of oxo-esters and thio-esters and to determine the rate-limiting step of catalysis for each substrate. The interest and limitation of this method is discussed. Molecular docking was used to analyze how the mutations could have altered the binding of the oxo-ester or the thio-ester. Results indicate that substitution of the ethereal oxygen for sulfur in substrates may alter the adjustment of substrate in the active site and stabilization of the transition-state for acylation. This affects the k2/k3 ratio and, in turn, controls the rate-limiting step of the hydrolytic reaction. Stabilization of the transition state is modulated both by the alcohol and acyl moieties of substrate. Interaction of these groups with the ethereal hetero-atom can have a neutral, an additive or an antagonistic effect on transition state stabilization, depending on their molecular structure, size and enantiomeric configuration.

Original languageEnglish (US)
Pages (from-to)16-34
Number of pages19
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1774
Issue number1
DOIs
StatePublished - Jan 1 2007

Fingerprint

Butyrylcholinesterase
Hydrolysis
Esters
Stabilization
Substrates
Mutation
Benzoylcholine
Acylation
Molecular Structure
Catalysis
Sulfur
Molecular structure
Cations
Catalytic Domain
Substitution reactions
Binding Sites
Alcohols
Oxygen
Atoms
Enzymes

Keywords

  • Butyrylcholinesterase
  • Ester
  • Mutation
  • Oxyanion hole
  • Rate-limiting step
  • Thio-ester

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Hydrolysis of oxo- and thio-esters by human butyrylcholinesterase. / Masson, Patrick; Froment, Marie Thérèse; Gillon, Emilie; Nachon, Florian; Lockridge, Oksana; Schopfer, Lawrence M.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1774, No. 1, 01.01.2007, p. 16-34.

Research output: Contribution to journalArticle

Masson, Patrick ; Froment, Marie Thérèse ; Gillon, Emilie ; Nachon, Florian ; Lockridge, Oksana ; Schopfer, Lawrence M. / Hydrolysis of oxo- and thio-esters by human butyrylcholinesterase. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2007 ; Vol. 1774, No. 1. pp. 16-34.
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