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 Citations (Scopus)

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 - 1982
Externally publishedYes

Fingerprint

Acetylcholinesterase
Acetylcholine
Hydrolysis
Phenolsulfonphthalein
Kinetics
Substrates
Acetylthiocholine
Edrophonium
Cholinesterase Inhibitors
Screening
Coloring Agents

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

A pseudo-first-order kinetic approach to measurement of acetylcholine hydrolysis by acetylcholinesterase. / Sharp, Thomas R.; Rosenberry, Terrone L.

In: Journal of Biochemical and Biophysical Methods, Vol. 6, No. 2, 1982, p. 159-172.

Research output: Contribution to journalArticle

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