Nucleophilic 1,4-additions for natural product discovery.
Nucleophilic 1,4-additions for natural product discovery.
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DOI：
10.1021/cb500324n
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发表时间：
2014-09-19
影响因子：
4
通讯作者：
Mitchell, Douglas A.
中科院分区：
文献类型：
--
作者：
Cox, Courtney L.;Tietz, Jonathan I.;Sokolowski, Karol;Melby, Joel O.;Doroghazi, James R.;Mitchell, Douglas A.
天然产品仍然是候选药物的重要来源，但传统分离固有的困难，加上化合物重新发现率高得令人无法接受，限制了天然产品检测的步伐。在这里，我们描述了一种基于反应性的筛选方法，以快速鉴定含有脱水氨基酸（即，羰基或亚胺活化的烯烃），这是几类天然产物中的常见基序。我们的策略需要使用市售的硫醇，二硫苏糖醇，通过亲核1，4-加成共价标记活化烯烃。通过比较反应和未反应的细胞表面提取物的质谱，可以很容易地辨别出修饰。当与推定的天然产物基因簇的生物信息学分析相结合时，可以对菌株的优先列表进行靶向筛选和分离。此外，已知的化合物很容易去复制，有效地消除了多余的分离和表征。作为原理的证明，该标记方法用于鉴定属于硫肽、羊毛硫肽和蒙花苷类的已知天然产物。此外，在筛选了一组只有23放线菌，我们发现并表征了一种新的硫肽抗生素，环噻唑霉素C。
Natural products remain an important source of drug candidates, but the difficulties inherent to traditional isolation, coupled with unacceptably high rates of compound rediscovery, limit the pace of natural product detection. Here we describe a reactivity-based screening method to rapidly identify exported bacterial metabolites that contain dehydrated amino acids (i.e., carbonyl- or imine-activated alkenes), a common motif in several classes of natural products. Our strategy entails the use of a commercially available thiol, dithiothreitol, for the covalent labeling of activated alkenes by nucleophilic 1,4-addition. Modification is easily discerned by comparing mass spectra of reacted and unreacted cell surface extracts. When combined with bioinformatic analysis of putative natural product gene clusters, targeted screening and isolation can be performed on a prioritized list of strains. Moreover, known compounds are easily dereplicated, effectively eliminating superfluous isolation and characterization. As a proof of principle, this labeling method was used to identify known natural products belonging to the thiopeptide, lanthipeptide, and linaridin classes. Further, upon screening a panel of only 23 actinomycetes, we discovered and characterized a novel thiopeptide antibiotic, cyclothiazomycin C.
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影响因子：
15
作者：
Bowers, Albert A.;Walsh, Christopher T.;Acker, Michael G.
通讯作者：
Acker, Michael G.
影响因子：
15
作者：
Dunbar, Kyle L.;Mitchell, Douglas A.
通讯作者：
Mitchell, Douglas A.
DOI：
10.1126/science.1176667
发表时间：
2009-08-28
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Fischbach MA;Walsh CT
通讯作者：
Walsh CT
影响因子：
1.8
作者：
AOKI, M;OHTSUKA, T;SETO, H
通讯作者：
SETO, H
影响因子：
1.8
作者：
AOKI, M;OHTSUKA, T;SETO, H
通讯作者：
SETO, H