A Biosynthetic Route to Photoclick Chemistry on Proteins

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• 作者：Jiangyun Wang ; Wei Zhang ; Wenjiao Song ; Yizhong Wang ; Zhipeng Yu ; Jiasong Li ; Minhao Wu ; Lin Wang ; Jianye Zang ; Qing Lin
• 刊名：Journal of the American Chemical Society
• 出版年：2010
• 出版时间：October 27, 2010
• 年：2010
• 卷：132
• 期：42
• 页码：14812-14818
• 全文大小：280K
• 年卷期：v.132,no.42(October 27, 2010)
• ISSN：1520-5126

文摘

Light-induced chemical reactions exist in nature, 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.