Evaluation of the decarbamylation process of cholinesterase during assay of enzyme activity.
The activity of carbamylated cholinesterase increases continuously during assay, suggesting that progressive decarbamylation takes place. The following effects of assay conditions on the observed decarbamylation were studied: the effect of the sulfhydryl group of nitrobenzoate produced in the course of Ellman assay, the effect of substrate and the effect of sample dilution during assay. This study indicates that sample dilution is the main trigger to the decarbamylation observed during assay of cholinesterase activity. The process was described as a first-order reaction during which the inhibited enzyme gives place to the active form. Kinetic constants for decarbamylation of human pseudocholinesterase (EC 3.1.1.8) at 30 degrees C were approximately 0.005 min-1 for dimethylcarbamates and 0.010 min-1 for monomethylcarbamates, when 1 mmol/l propionylthiocholine was used as substrate.
Institute of Clinical Toxicology and Pharmacology, Sheba Medical Center, Tel Hashomer, Israel.
M
M
Rotenberg
S
S
Almog
D000448:Aldicarb / Q000032:analysis / Q000097:blood / Q000378:metabolism; D002219:Carbamates / Q000032:analysis / Q000097:blood / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D002801:Cholinesterase Reactivators / Q000494:pharmacology; D002802:Cholinesterases / Q000032:analysis / Q000097:blood / Q000378:metabolism*; D006801:Humans; D007201:Indicator Dilution Techniques; D007306:Insecticides / Q000032:analysis / Q000097:blood / Q000378:metabolism; D007700:Kinetics; D011729:Pyridostigmine Bromide / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000494:pharmacology
D002219:Carbamates; D002800:Cholinesterase Inhibitors; D002801:Cholinesterase Reactivators; D007306:Insecticides; D013438:Sulfhydryl Compounds; D000448:Aldicarb; D002802:Cholinesterases; D011729:Pyridostigmine Bromide