Studies of reversible inhibition, irreversible inhibition, and activation of alkaline phosphatase by capillary electrophoresis
Reversible inhibition, irreversible inhibition, and activation of calf intestinal alkaline phosphatase (EC 126.96.36.199) have been studied by capillary electrophoresis. The capillary electrophoretic enzyme-inhibitor assays were based on electrophoretic mixing of inhibitor and enzyme zones in a substrate-filled capillary. Enzyme inhibition was indicated by a decrease in product formation detected in the capillary by laser-induced fluorescence. Reversible enzyme inhibitors could be quantified by Michaelis-Menten treatment of the electrophoretic data. Reversible, competitive inhibition of alkaline phosphatase by sodium vanadate and sodium arsenate has been examined, and reversible, noncompetitive inhibition by theophylline has been studied. The Ki values determined for these reversible inhibitors using capillary electrophoresis are within the range of values reported in the literature for the same enzyme-inhibitor combinations. Irreversible inhibition of alkaline phosphatase by EDTA at concentrations of 1.0 mM and above has been observed. Activation of alkaline phosphatase has also been observed for EDTA at concentrations from 20 to 400 μM. © 2002 Elsevier Science (USA). All rights reserved.
Publication Source (Journal or Book title)
Whisnant, A., & Gilman, S. (2002). Studies of reversible inhibition, irreversible inhibition, and activation of alkaline phosphatase by capillary electrophoresis. Analytical Biochemistry, 307 (2), 226-234. https://doi.org/10.1016/S0003-2697(02)00062-3