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Anal Biochem ; 343(2): 293-8, 2005 Aug 15.
Article in English | MEDLINE | ID: mdl-16004952

ABSTRACT

In this work, we report an alternative assay for the determination of the inhibitory effect on monoamine oxidase B (MAO-B) activity of probe compounds. Enzyme MAO-B exhibits fluorescence emissions when it is excited at 412 nm. Using an inexpensive blue LED-like excitation source, we measured the quenching of fluorescence intensity of MAO-B enzyme during the reaction with inhibitors. The applicability of the procedure is demonstrated by assays with l-deprenyl and berberine as inhibitors through the use of fluorescence studies. The IC(50) values of l-deprenyl and berberine were 0.04 and 90 microM, respectively. The K(I) values were 0.020 and 47 microM for l-deprenyl and berberine, respectively. These IC(50) and K(I) values were similar to the values obtained with a standard method. These results demonstrate the feasibility of this method as an alternative to follow the inhibitory effect on MAO-B.


Subject(s)
Monoamine Oxidase/metabolism , Spectrometry, Fluorescence/methods , Animals , Benzylamines/chemistry , Berberine/chemistry , Berberine/pharmacology , Dose-Response Relationship, Drug , Kinetics , Light , Mice , Mitochondria, Liver/drug effects , Mitochondria, Liver/metabolism , Monoamine Oxidase Inhibitors/metabolism , Selegiline/chemistry , Selegiline/pharmacology
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