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基于高效液相色谱的植物提取物生物活性分析：一种使用人重组单胺氧化酶-A鉴定单胺氧化酶-A抑制剂的动力学测定法。

HPLC-based bioactivity profiling of plant extracts: a kinetic assay for the identification of monoamine oxidase-A inhibitors using human recombinant monoamine oxidase-A.

作者信息

Dittmann Kathrin, Riese Ulrike, Hamburger Matthias

机构信息

Institute of Pharmacy, Friedrich-Schiller-University Jena, Semmelweisstrasse 10, D-07743 Jena, Germany.

出版信息

Phytochemistry. 2004 Nov;65(21):2885-91. doi: 10.1016/j.phytochem.2004.07.024.

DOI:10.1016/j.phytochem.2004.07.024
PMID:15501256
Abstract

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.

摘要

建立了一种基于高效液相色谱法在植物提取物中寻找单胺氧化酶-A（MAO-A）抑制剂的检测方法。该方法将在酵母中表达为谷胱甘肽S-转移酶（GST）融合蛋白的人重组MAO-A与犬尿胺转化为4-羟基喹啉的动力学测定相结合。GST融合蛋白的底物选择性和动力学参数与野生型酶相当。用添加少量已知MAO抑制剂的植物提取物测试了该检测方法在基于高效液相色谱法的活性分析中的适用性。

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