Targeting Reactive Carbonyls for Identifying Natural Products and Their Biosynthetic Origins.
Targeting Reactive Carbonyls for Identifying Natural Products and Their Biosynthetic Origins.
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DOI：
10.1021/jacs.6b06848
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发表时间：
2016-11-23
影响因子：
15
通讯作者：
Mitchell DA
中科院分区：
文献类型：
--
作者：
Maxson T;Tietz JI;Hudson GA;Guo XR;Tai HC;Mitchell DA
天然产物作为候选药物和化学生物学的工具具有重要的作用。然而，传统的天然产物发现，主要基于生物测定指导的方法，偏向于丰富的化合物和重新发现率高。因此，需要正交方法来促进新的天然产物的发现，在这里，我们描述了基于同位素标记的基于反应性的天然产物筛选的扩展，以解决这些缺点。基于反应性的筛选是一种定向发现方法，其中化学选择探针针对天然产物的特定反应处理，使其能够通过质谱检测。在这项研究中，我们开发了一种含氨基的探针，以指导发现含醛和酮的天然产物。为了便于检测标记事件，探针被二溴化，赋予独特的同位素特征，以区分标记的代谢物和光谱噪声。作为概念的证明，该探针随后被用于筛选细菌提取物的集合，从而鉴定出一种新的抗痛类似物，deimino-antipain。通过生物信息学分析和异源表达鉴定了产生deimino-antipain的细菌序列，并鉴定了相关的生物合成基因簇。这些数据揭示了一个众所周知的含有醛的肽类蛋白酶抑制剂家族的未知遗传基础，包括止痛药、凝乳素、白细胞蛋白酶、弹性蛋白酶和微生物碱性蛋白酶抑制剂（MAPI），这些抑制剂已被广泛使用了40多年。
Natural products serve important roles as drug candidates and as tools for chemical biology. However, traditional natural product discovery, largely based on bioassay-guided approaches, is biased towards abundant compounds and rediscovery rates are high. Orthogonal methods to facilitate discovery of new natural products are thus needed, and herein we describe an isotope tag-based expansion of reactivity-based natural product screening to address these shortcomings. Reactivity-based screening is a directed discovery approach in which a specific reactive handle on the natural product is targeted by a chemoselective probe to enable its detection by mass spectrometry. In this study, we have developed an aminooxy-containing probe to guide the discovery of aldehyde- and ketone-containing natural products. To facilitate the detection of labeling events, the probe was dibrominated, imparting a unique isotopic signature to distinguish labeled metabolites from spectral noise. As a proof of concept, the probe was then utilized to screen a collection of bacterial extracts, leading to the identification of a new analog of antipain, deimino-antipain. The bacterial producer of deimino-antipain was sequenced and the responsible biosynthetic gene cluster was identified by bioinformatic analysis and heterologous expression. These data reveal the previously undetermined genetic basis for a well-known family of aldehyde-containing, peptidic protease inhibitors, including antipain, chymostatin, leupeptin, elastatinal, and microbial alkaline protease inhibitor (MAPI), which have been widely used for over 40 years.
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影响因子：
2
作者：
HOEBEKE, J;BUSATTOSAMSOEN, C;LEBRUN, E
通讯作者：
LEBRUN, E
影响因子：
16.6
作者：
通讯作者：
--
影响因子：
14.9
作者：
Clark K;Karsch-Mizrachi I;Lipman DJ;Ostell J;Sayers EW
通讯作者：
Sayers EW
影响因子：
4
作者：
Castro-Falcón G;Hahn D;Reimer D;Hughes CC
通讯作者：
Hughes CC
影响因子：
4.4
作者：
Felnagle, Elizabeth A.;Rondon, Michelle R.;Thomas, Michael G.
通讯作者：
Thomas, Michael G.