HPLC-based bioactivity profiling of plant extracts: a kinetic assay for the identification of monoamine oxidase-A inhibitors using human recombinant monoamine oxidase-A.
An assay for the HPLC-based search for monoamine oxidase-A (MAO-A) inhibitors in plant extracts was established. It combines human recombinant MAO-A, expressed as GST-fusion protein in yeast, with a kinetic measurement of the conversion of kynuramine to 4-hydroxyquinoline. Substrate selectivity and kinetic parameters of the GST-fusion protein were comparable to the wild-type enzyme. The applicability of the assay to HPLC-based activity profiling was tested with plant extracts spiked with small amounts of known MAO inhibitors.
Dittmann
Kathrin
K
Institute of Pharmacy, Friedrich-Schiller-University Jena, Semmelweisstrasse 10, D-07743 Jena, Germany.
Riese
Ulrike
U
Hamburger
Matthias
M
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007700:Kinetics; D008995:Monoamine Oxidase / Q000378:metabolism*; D008996:Monoamine Oxidase Inhibitors / Q000494:pharmacology*; D010936:Plant Extracts / Q000737:chemistry* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D012680:Sensitivity and Specificity; D013379:Substrate Specificity
D008996:Monoamine Oxidase Inhibitors; D010936:Plant Extracts; D011994:Recombinant Proteins; D008995:Monoamine Oxidase