A biosynthetic route to photoclick chemistry on proteins.

A biosynthetic route to photoclick chemistry on proteins.
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DOI:
10.1021/ja104350y
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发表时间:
2010-10-27
影响因子:
15
通讯作者:
Lin, Qing
Lin, Qing
中科院分区:
化学1区
文献类型:
--
作者:
Wang, Jiangyun;Zhang, Wei;Song, Wenjiao;Wang, Yizhong;Yu, Zhipeng;Li, Jiasong;Wu, Minhao;Wang, Lin;Zang, Jianye;Lin, Qing

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光诱导的化学反应存在于自然界中,调节许多重要的细胞和生物体功能,例如,原核生物的感光和哺乳动物的视觉形成。在这里,我们报告了光反应性非天然氨基酸p-(2-四唑)苯丙氨酸(p-Tpa)基因整合到大肠杆菌中的肌红蛋白位点特异性。大肠杆菌通过进化一个正交的tRNA/氨酰-tRNA合成酶对,和使用p-Tpa作为生物正交的化学“手柄”的荧光标记的p-Tpa编码的肌红蛋白通过photoclick反应。此外,我们阐明的结构基础的p-Tpa的生物合成掺入到蛋白质中,通过解决的X射线结构的p-Tpa-特异性氨酰-tRNA合成酶与p-Tpa的复合物。这种光反应性氨基酸的遗传编码应该使未来有可能在生命系统中光调节蛋白质功能。
Light-induced chemical reactions exist in nature in regulating many important cellular and organismal functions, e.g., photosensing in prokaryotes and vision formation in mammals. Here, we report the genetic incorporation of a photoreactive unnatural amino acid, p-(2-tetrazole)phenylalanine (p-Tpa), into myoglobin site-specifically in E. coli by evolving an orthogonal tRNA/aminoacyl-tRNA synthetase pair, and the use of p-Tpa as a bioorthogonal chemical “handle” for fluorescent labeling of p-Tpa-encoded myoglobin via the photoclick reaction. Moreover, we elucidated the structural basis for the biosynthetic incorporation of p-Tpa into proteins by solving the X-ray structure of p-Tpa-specific aminoacyl-tRNA synthetase in complex with p-Tpa. The genetic encoding of this photoreactive amino acid should make it possible in the future to photoregulate protein function in living systems.
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