One-Pot Synthesis of Azoline-Containing Peptides in a Cell-free Translation System Integrated with a Posttranslational Cyclodehydratase
One-Pot Synthesis of Azoline-Containing Peptides in a Cell-free Translation System Integrated with a Posttranslational Cyclodehydratase
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DOI：
10.1016/j.chembiol.2014.04.008
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发表时间：
2014-06-19
影响因子：
--
通讯作者：
Suga, Hiroaki
中科院分区：
文献类型：
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作者：
Goto, Yuki;Ito, Yumi;Suga, Hiroaki
多肽类天然产物骨架中的唑啉基团是具有多种生物活性的重要结构基元。一些含氮唑啉的多肽(Az-多肽)是由核糖体合成的前体多肽产生的，其中半胱氨酸、丝氨酸和苏氨酸残基通过环脱水酶的翻译后修饰转化为相应的唑类。我们通过将无细胞翻译系统与翻译后环状脱水酶PatD相结合，设计了一种Az多肽的体外生物合成系统，称为FIT-PatD(柔性体外翻译)系统。该系统能够“一锅法”合成从合成DNA模板表达的多种Az多肽衍生物。Fit-PatD系统还促进了对一系列前体多肽序列的突变研究，揭示了PatD独特的体外底物耐受性。
Azoline moieties in the backbones of peptidic natural products are important structural motifs that contribute to diverse bioactivities. Some azoline-containing peptides (Az-peptides) are produced from ribosomally synthesized precursor peptides, in which cysteine, serine, and threonine residues are converted to their corresponding azolines by post-translational modification through a cyclodehydratase. We have devised an in vitro biosynthesis system of Az-peptides, referred to as the FIT-PatD (flexible in vitro translation) system, by the integration of a cell-free translation system with the post-translational cyclodehydratase PatD. This system enabled the "one-pot'' synthesis of a wide variety of Az-peptide derivatives expressed from synthetic DNA templates. The FIT-PatD system also facilitated mutagenesis studies on a wide array of precursor peptide sequences, unveiling unique in vitro substrate tolerance of PatD.