Luo Xiaozhou, Zambaldo Claudio, Liu Tao, Zhang Yuhan, Xuan Weimin, Wang Chen, Reed Sean A, Yang Peng-Yu, Wang Rongsheng E, Javahishvili Tsotne, Schultz Peter G, Young Travis S
Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037;
Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037; Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037; Department of Biology, California Institute for Biomedical Research, La Jolla, CA 92037.
Proc Natl Acad Sci U S A. 2016 Mar 29;113(13):3615-20. doi: 10.1073/pnas.1602733113. Epub 2016 Mar 14.
Thiopeptides are a subclass of ribosomally synthesized and posttranslationally modified peptides (RiPPs) with complex molecular architectures and an array of biological activities, including potent antimicrobial activity. Here we report the generation of thiopeptides containing noncanonical amino acids (ncAAs) by introducing orthogonal amber suppressor aminoacyl-tRNA synthetase/tRNA pairs into a thiocillin producer strain of Bacillus cereus .We demonstrate that thiopeptide variants containing ncAAs with bioorthogonal chemical reactivity can be further postbiosynthetically modified with biophysical probes, including fluorophores and photo-cross-linkers. This work allows the site-specific incorporation of ncAAs into thiopeptides to increase their structural diversity and probe their biological activity; similar approaches can likely be applied to other classes of RiPPs.
硫肽是核糖体合成并经翻译后修饰的肽(RiPPs)的一个亚类,具有复杂的分子结构和一系列生物活性,包括强大的抗菌活性。在此,我们报告通过将正交琥珀抑制型氨酰-tRNA合成酶/tRNA对引入蜡状芽孢杆菌的硫代青霉素生产菌株中,生成含有非天然氨基酸(ncAAs)的硫肽。我们证明,含有具有生物正交化学反应性的非天然氨基酸的硫肽变体可以用包括荧光团和光交联剂在内的生物物理探针进行进一步的生物合成后修饰。这项工作允许将非天然氨基酸位点特异性地掺入硫肽中,以增加其结构多样性并探究其生物活性;类似的方法可能也适用于其他类别的RiPPs。