1. Inhibition kinetics of human serum butyrylcholinesterase by Cd2+, Zn2+ and Al3+: comparison of the effects of metal ions on cholinesterases
- Author
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E. Ferhan Tezcan, Bahram Sarkarati, and A. Neşe Çokuğraş
- Subjects
Metal ions in aqueous solution ,Immunology ,Allosteric regulation ,Butyrylthiocholine ,Enzyme Reactivators ,Affinity chromatography ,Humans ,Magnesium ,Butyrylcholinesterase ,Pharmacology ,chemistry.chemical_classification ,Chromatography ,biology ,Cooperative binding ,Enzyme assay ,Kinetics ,Zinc ,Enzyme ,chemistry ,biology.protein ,Calcium ,Cholinesterase Inhibitors ,Allosteric Site ,Aluminum ,Cadmium - Abstract
Butyrylcholinesterase (BChE, EC 3.1.1.8) has been purified about 6600-fold from human serum with a procedure including ammonium sulfate fractionation (55–70%) with acid step at pH 4.5 and procainamide–Sepharose 4B affinity chromatography. The purified enzyme exhibited negative cooperativity with respect to butyrylthiocholine (BTCh) binding at pH 7.5. KS was found to be 0.128±0.012 mM. Inhibition kinetics of the enzyme by Cd2+, Zn2+ and Al3+ were studied in detail. The 1/v vs 1/[BTCh] plots in the absence (control plot) and in the presence of different concentrations of cations intersected above 1/[BTCh]-axis. The data were analyzed by means of a nonlinear curve fitting program. The results demonstrated that all of the three cations are the linear mixed-type inhibitors of BChE. Ca2+ and Mg2+ had no effect on the enzyme activity in the experimental conditions. But when the enzyme was inhibited by 0.5 mM Cd2+ or Zn2+, Ca2+ and Mg2+ partially reactivated the inhibited allosteric form of BChE. Results were compared with data obtained from brain BChE purified from sheep.
- Published
- 1999
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