TY - JOUR
T1 - A pseudo-first-order kinetic approach to measurement of acetylcholine hydrolysis by acetylcholinesterase
AU - Sharp, Thomas R.
AU - Rosenberry, Terrone L.
N1 - Funding Information:
T.R.S. is a Postdoctoral Research Fellow of the Muscular Dystrophy Association of America. Financial support for this study was provided by National Institutes of
PY - 1982/6
Y1 - 1982/6
N2 - 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+.
AB - 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+.
KW - acetylcholinesterase
KW - aluminum inhibition
KW - decamethonium inhibition
KW - edrophonium inhibition
KW - first-order kinetics method
KW - inhibition constants
KW - phenol red
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U2 - 10.1016/0165-022X(82)90062-8
DO - 10.1016/0165-022X(82)90062-8
M3 - Article
C2 - 7108127
AN - SCOPUS:0020149579
SN - 0165-022X
VL - 6
SP - 159
EP - 172
JO - Journal of Biochemical and Biophysical Methods
JF - Journal of Biochemical and Biophysical Methods
IS - 2
ER -