In Vitro Biosynthesis of Peptides Containing Exotic Azoline Analogues
In Vitro Biosynthesis of Peptides Containing Exotic Azoline Analogues
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DOI：
10.1002/cbic.201900521
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发表时间：
2019-11-18
期刊：
影响因子：
3.2
通讯作者：
Suga, Hiroaki
中科院分区：
文献类型：
--
作者：
Goto, Yuki;Suga, Hiroaki
在自然界中，在核糖体合成和后修饰的肽（RIPPs）的生物合成期间由YcaO环化酶产生的唑啉通常限于噻唑啉、恶唑啉和甲基恶唑啉，其分别衍生自蛋白原Cys、Ser和Thr残基。为了研究YcaO环化酶是否精确识别可修饰残基的共同结构特征和手性，通过将YcaO环化酶（PatD）与由柔性体外翻译（FIT）系统提供动力的遗传密码重编程相结合，建立了“重编程FIT-PatD系统”，其中带有非蛋白质生成Cys/Ser/Thr类似物的前体肽可以通过重编程遗传密码表达，随后通过PatD环脱水。这项研究揭示了显着的立体，化学，和regioversatility的可修饰的残基在PatD催化的环化脱水，扩大剧目的骨干杂环RIPPs外来的唑啉类似物。
In nature, azolines produced by YcaO cyclodehydratases during the biosynthesis of ribosomally synthesized and post-translationally modified peptides (RiPPs) are generally limited to thiazoline, oxazoline, and methyloxazoline, which are derived from the proteinogenic Cys, Ser, and Thr residues, respectively. To investigate whether YcaO cyclodehydratases precisely recognize the common structural characteristics and chirality of the modifiable residues, the "reprogrammed FIT-PatD system" has been established by combining a YcaO cyclodehydratase (PatD) with genetic code reprogramming powered by the flexible in vitro translation (FIT) system, in which precursor peptides bearing non-proteinogenic Cys/Ser/Thr analogues could be expressed through a reprogrammed genetic code and subsequently cyclodehydrated by PatD. The study has revealed remarkable stereo-, chemo-, and regioversatility for modifiable residues in PatD-catalyzed cyclodehydration, expanding the repertoire of backbone heterocycles in RiPPs to exotic azoline analogues.