Zhou Wenhui, Valley Michael P, Shultz John, Hawkins Erika M, Bernad Laurent, Good Troy, Good Dave, Riss Terry L, Klaubert Dieter H, Wood Keith V
Promega Biosciences, Inc., 277 Granada Drive, San Luis Obispo, California 93401, USA.
J Am Chem Soc. 2006 Mar 15;128(10):3122-3. doi: 10.1021/ja058519o.
Novel bioluminogenic substrates were designed for probing monoamine oxidase (MAO) activity based on a simple and effective beta-elimination strategy. By modifying the amino group and the central core of luciferin derivatives, we have developed a series of substrates useful for assays of MAO A or B, or both. One of these substrates, exhibiting low Km values and high signal-to-background ratios with both isozymes, was shown to accurately measure the Ki values of known MAO inhibitors. This substrate is a key component in the development of a highly sensitive homogeneous MAO assay for high-throughput screening (HTS) of compounds in drug discovery and for monitoring MAO activity in complex biological systems. This design strategy should be applicable to fluorogenic MAO substrates and could broaden the structural requirements of substrates for other enzyme assays.
基于一种简单有效的β-消除策略，设计了新型生物发光底物用于探测单胺氧化酶（MAO）活性。通过修饰荧光素衍生物的氨基和核心结构，我们开发了一系列可用于检测MAO A或B或两者的底物。其中一种底物对两种同工酶均表现出低Km值和高信噪比，已证明其能准确测量已知MAO抑制剂的Ki值。该底物是开发高灵敏度均相MAO检测方法的关键组成部分，可用于药物发现中化合物的高通量筛选（HTS）以及监测复杂生物系统中的MAO活性。这种设计策略应适用于荧光MAO底物，并可拓宽其他酶检测底物的结构要求。
