PMID: 11916145Mar 28, 2002Paper

Progress curves analysis as an alternative for exploration of activation-inhibition phenomena in cholinesterases

Journal of Enzyme Inhibition
M Golicnik

Abstract

The kinetic behaviour of insect acetylcholinesterases deviates from the Michaelis-Menten pattern. These deviations are known as activation or inhibition at various substrate concentrations and can be more or less observable depending on mutations around the active site of the enzyme. Most kinetic studies on these enzymes still rely on initial rate measurements. It is demonstrated here that according to this method one of the deviations can be overlooked. We attempt to point out that in such cases a detailed step-by-step progress curves analysis is successful. The study is focused on two different methods of analysing progress curves: (i) the first one is based on an integrated initial rate equation which can sufficiently fit truncated progress curves under corresponding conditions; and (ii) the other one precludes the algebraic formulae, but uses numerical integration for searching a non analytical solution of ordinary differential equations describing a kinetic model. All methods are tested on three different acetylcholinesterase mutants from Drosophila melanogaster. The results indicate that kinetic parameters for the E107K mutant with highly expressive activation and inhibition can be well evaluated applying any analysis met...Continue Reading

References

Jun 1, 1987·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·G CauetD Thomas
Jun 1, 1973·Canadian Journal of Biochemistry·N H Hijazi, K J Laidler
Mar 31, 1998·Protein Expression and Purification·S Estrada-Mondaca, D Fournier
Jul 1, 1961·Biochemical Pharmacology·G L ELLMANR M FEATHER-STONE

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