Multitarget affinity/specificity screening of natural products: Finding and characterizing high-affinity ligands from complex mixtures by using high-performance mass spectrometry
Multitarget affinity/specificity screening of natural products: Finding and characterizing high-affinity ligands from complex mixtures by using high-performance mass spectrometry
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DOI：
10.1021/np0301137
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发表时间：
2003-09-01
影响因子：
5.1
通讯作者：
Hofstadler, SA
中科院分区：
文献类型：
--
作者：
Cummins, LL;Chen, S;Hofstadler, SA
在这项工作中，我们描述了一种基于电喷雾电离-傅里叶变换离子回旋共振质谱 (ESI-FTICR) 的高通量筛选方法，该方法可快速询问基于 RNA 的药物靶标与源自细菌天然产物库的成分之间的非共价相互作用。筛选过程检测天然产物文库中存在的分子，这些分子与模拟原核16S rRNA A位点的合成RNA靶标结合，同时使用缺乏A位点构建体关键结构元件的对照RNA靶标测量合成A位点靶标的特异性。这种称为多靶点亲和力/特异性筛选 (MASS) 的筛选方法证明了巴龙霉素与来自裂缘链霉菌 (Streptomyces rimosus sp.) 的分级天然产物文库的预期结合。巴龙霉素。观察到一种新分子与 16S A 位点 RNA 构建体特异性结合。该物种的 MS/MS 表征产生了部分结构信息，表明它是一种氨基糖苷，由具有一个或多个修饰环的巴龙霉素核心组成。这项工作展示了 MASS 在针对大分子靶点筛选天然产物组分方面的巨大效用。
In this work we describe a high-throughput screening approach based on electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR) that rapidly interrogates the noncovalent interaction between RNA-based drug targets and components derived from a bacterial natural product library. The screening process detects molecules present in the natural product library that bind to a synthetic RNA target that mimics the prokaryotic 16S rRNA A-site, while simultaneously measuring specificity for the synthetic A-site target using a control RNA target that lacks the critical structural element of the A-site construct. This screening approach known as multitarget affinity/specificity screening (MASS) demonstrated the expected binding of paromomycin from a fractionated natural product library derived from Streptomyces rimosus sp. paromomycinus. A new molecule was observed to bind with specificity to the 16S A-site RNA construct. MS/MS characterization of this species yielded partial structural information suggesting it is an aminoglycoside consisting of a paromomycin core with one or more modified rings. This work demonstrates the tremendous utility of MASS for screening natural product fractions against macromolecular targets.