COMPUTERIZED ANALYSIS OF COVALENT INHIBITION-KINETICS FOR IDENTIFICATION OF HEART-MUSCLE CHOLINESTERASE AND BRAIN CARBOXYLESTERASE ISOENZYMES - DESIGN OF DIFFERENTIAL INHIBITION ASSAYS

The kinetics of time- and concentration-dependent covalent organophosphorus inhibition of carboxylesterase isoenzymes (EC 3.1.1.1) and cholinesterase isoenzymes (EC 3.1.1.7 and EC 3.1.1.8) were investigated using a wide range of organophosphate inhibitor concentrations (10(-10)- 10(-3) mol/l) and different inhibition times. Computerized analysis of inhibition curves by weighted nonlinear least-squares curve fitting was compared to graphic analysis by iterative elimination of exponential functions. Possible experimental errors due to inhibitor saturation kinetics and enzymatic organophosphate hydrolysis were thoroughly investigated. In mammalian heart muscle, three different cholinesterase isoenzymes were identified. High sensitivity and specificity of the classic differential inhibition test for carboxylesterase activity of hen brain neuropathy target esterase (NTE) could be confirmed independently with both methods of inhibition curve analysis.