Selective activity of butyrylcholinesterase in serum by a chemiluminescent assay.
In a previous study, we showed that purified commercial esterase activity can be detected in a chemiluminescent assay based on the hydrolysis of 2-methyl-1-propenylbenzoate (MPB) to 2-methyl-1-propenol, which is subsequently oxidized by the horseradish peroxidase (HRP)-H(2)O(2) system. The purpose of this study was to verify the applicability of this assay to human serum. The existence of an esterase activity capable of hydrolysing MPB is indicated by the fact that the MPB-serum-HRP-H(2)O(2) system consumes oxygen and emits light. Both signals were abolished by prior serum heat inactivation and were preserved when serum was stored at < or =4 degrees C. Addition of aliesterase inhibitors, such as fluoride ion and trichlorfon or the cholinesterase inhibitor eserine, totally prevents light emission. The butyrylcholinesterase-specific substrate benzoylcholine causes a delay in both O(2) uptake and light emission, while the specific acetylcholinesterase substrate, acetyl-beta-methylcholine, had practically no effect. Purified butyrylcholinesterase, but not acetylcholinesterase, triggered light emission. The finding that butyrylcholinesterase is responsible for the hydrolysis of MPB in serum should serve as the basis for the development of a specific chemiluminescent assay for this enzyme.
Yavo
B
B
Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, CP 66.083, 05389-970 São Paulo, Brazil.
Brunetti
I L
IL
da Fonseca
L M
LM
Catalani
L H
LH
Campa
A
A
D000818:Animals; D001565:Benzoates / Q000737:chemistry* / Q000378:metabolism; D001588:Benzoylcholine / Q000737:chemistry / Q000378:metabolism; D000440:Butanols / Q000737:chemistry* / Q000378:metabolism; D002091:Butyrylcholinesterase / Q000097:blood*; D002417:Cattle; D002800:Cholinesterase Inhibitors / Q000737:chemistry* / Q000378:metabolism; D004912:Erythrocytes / Q000201:enzymology; D005459:Fluorides / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008163:Luminescent Measurements; D016210:Methacholine Chloride / Q000737:chemistry / Q000378:metabolism; D010830:Physostigmine / Q000737:chemistry / Q000378:metabolism; D014236:Trichlorfon / Q000737:chemistry
C101261:2-methyl-1-propenylbenzoate; D001565:Benzoates; D000440:Butanols; D002800:Cholinesterase Inhibitors; D016210:Methacholine Chloride; D001588:Benzoylcholine; C040507:isobutyl alcohol; D010830:Physostigmine; D014236:Trichlorfon; D006735:Horseradish Peroxidase; D002091:Butyrylcholinesterase; D005459:Fluorides