Shang X, Song X, Faller C, Lai R, Li H, Cerny R, Niu W, Guo J
Department of Chemistry , University of Nebraska-Lincoln , Lincoln , NE 68588 , USA . Email:
Department of Chemical & Biomolecular Engineering , University of Nebraska-Lincoln , Lincoln , NE 68588 , USA . Email:
Chem Sci. 2017 Feb 1;8(2):1141-1145. doi: 10.1039/c6sc03635j. Epub 2016 Sep 26.
We developed a new fluorogenic bioorthogonal reaction that is based on the inverse electron-demand Diels-Alder reaction between styrene (an unstrained alkene) and a simple tetrazine. The reaction forms a new fluorophore with no literature precedent. We have identified an aminoacyl-tRNA synthetase/tRNA pair for the efficient and site-specific incorporation of a styrene-containing amino acid into proteins in response to amber nonsense codon. Fluorogenic labeling of purified proteins and intact proteins in live cells were demonstrated. The fluorogenicity of the styrene-tetrazine reaction can be potentially applied to the study of protein folding and function under physiological conditions with low background fluorescence interference.
我们开发了一种新的荧光生物正交反应，该反应基于苯乙烯（一种非张力烯烃）与一种简单四嗪之间的逆电子需求狄尔斯-阿尔德反应。该反应形成了一种无文献先例的新荧光团。我们已经鉴定出一种氨酰-tRNA合成酶/tRNA对，用于在响应琥珀色无义密码子时将含苯乙烯的氨基酸高效且位点特异性地掺入蛋白质中。展示了对纯化蛋白质和活细胞中完整蛋白质的荧光标记。苯乙烯-四嗪反应的荧光性有可能应用于在低背景荧光干扰的生理条件下研究蛋白质折叠和功能。
