Excited-state intermediates in a designer protein encoding a phototrigger caught by an X-ray free-electron laser
Excited-state intermediates in a designer protein encoding a phototrigger caught by an X-ray free-electron laser
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DOI：
10.1038/s41557-022-00992-3
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发表时间：
2022-07
期刊：
影响因子：
21.8
通讯作者：
Xiaohong Liu;Pengcheng Liu;Hongjie Li;Zhen Xu;Lu Jia;Yangsheng Xia;Minling Yu;Wen-Yi Tang;Xiaolei Zhu;Chao Chen;Yuanlin Zhang;E. Nango;Rie Tanaka;F. Luo;Koji Kato;Y. Nakajima;Shunpei Kishi;Huaxin Yu;N. Matsubara;S. Owada;K. Tono;S. Iwata;Long-Jiang Yu;Jian-Ren Shen;Jiangyun Wang
中科院分区：
文献类型：
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作者：
Xiaohong Liu;Pengcheng Liu;Hongjie Li;Zhen Xu;Lu Jia;Yangsheng Xia;Minling Yu;Wen-Yi Tang;Xiaolei Zhu;Chao Chen;Yuanlin Zhang;E. Nango;Rie Tanaka;F. Luo;Koji Kato;Y. Nakajima;Shunpei Kishi;Huaxin Yu;N. Matsubara;S. Owada;K. Tono;S. Iwata;Long-Jiang Yu;Jian-Ren Shen;Jiangyun Wang
化学研究的主要目标之一是真实的观察反应过程中原子的运动。尽管X射线自由电子激光（XFEL）促进了使用时间分辨连续飞秒晶体学（TR-SFX）捕获反应中间体，但只有少数天然光敏蛋白质使用这种方法进行了研究，主要是由于缺乏合适的光触发剂。在这里，我们报告的遗传编码的氧杂蒽酮氨基酸（FXO），作为一个有效的光触发，到一个合理设计的人肝脂肪酸结合蛋白突变体（称为XOM），它经历了光诱导的C-H键转换具有高选择性和量子效率。我们以1.55-1.70 Å的分辨率解决了闪光照射前和闪光照射后10-300 ns的XOM结构，并捕获了负责精确C-H键激活的难以捉摸的激发态中间体。我们希望大多数氧化还原酶现在可以通过TR-SFX进行研究，使用我们的方法，以揭示其效率和选择性的关键反应中间体。
One of the primary objectives in chemistry research is to observe atomic motions during reactions in real time. Although X-ray free-electron lasers (XFELs) have facilitated the capture of reaction intermediates using time-resolved serial femtosecond crystallography (TR-SFX), only a few natural photoactive proteins have been investigated using this method, mostly due to the lack of suitable phototriggers. Here we report the genetic encoding of a xanthone amino acid (FXO), as an efficient phototrigger, into a rationally designed human liver fatty-acid binding protein mutant (termed XOM), which undergoes photo-induced C–H bond transformation with high selectivity and quantum efficiency. We solved the structures of XOM before and 10–300 ns after flash illumination, at 1.55–1.70 Å resolutions, and captured the elusive excited-state intermediates responsible for precise C–H bond activation. We expect that most redox enzymes can now be investigated by TR-SFX, using our method, to reveal reaction intermediates key for their efficiency and selectivity.