A pseudo-first-order kinetic approach to measurement of acetylcholine hydrolysis by acetylcholinesterase

Thomas R. Sharp, Terrone L. Rosenberry

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A continuous spectrophotometric procedure is presented for the measurement of the kinetic properties of acetylcholinesterase (EC 3.1.1.7) with its natural substrate, acetylcholine. The procedure is based upon the production of stoichiometric quantities of H+ upon hydrolysis of substrate. The spectrophotometric reporter is the pH indicator dye, phenol red and the procedure yields continuous time courses for hydrolysis of substrate. Further, this phenol red system and an adaptation of the Ellman et al. (1961, Biochem. Pharmacol. 7, 88-95) procedure for acetylthiocholine as substrate, are described as a rapid screening technique for reversible competitive and noncompetitive inhibitors of acetylcholinesterase activity. The methods are illustrated by determinations of K1 for edrophonium, decamethonium and Al3+.

Original languageEnglish (US)
Pages (from-to)159-172
Number of pages14
JournalJournal of Biochemical and Biophysical Methods
Volume6
Issue number2
DOIs
StatePublished - Jun 1982

Keywords

  • acetylcholinesterase
  • aluminum inhibition
  • decamethonium inhibition
  • edrophonium inhibition
  • first-order kinetics method
  • inhibition constants
  • phenol red

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

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