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Site-Specific Incorporation of a Thioester Containing Amino Acid into Proteins.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2018-02-09 , DOI: 10.1021/acschembio.7b00998
Weimin Xuan 1 , Daniel Collins 2 , Minseob Koh 1 , Sida Shao 1 , Anzhi Yao 1 , Han Xiao 1 , Philip Garner 2 , Peter G Schultz 1
Affiliation  

Here, we report the site-specific incorporation of a thioester containing noncanonical amino acid (ncAA) into recombinantly expressed proteins. Specifically, we genetically encoded a thioester-activated aspartic acid (ThioD) in bacteria in good yield and with high fidelity using an orthogonal nonsense suppressor tRNA/aminoacyl-tRNA synthetase (aaRS) pair. To demonstrate the utility of ThioD, we used native chemical ligation to label green fluorescent protein with a fluorophore in good yield.

中文翻译:

蛋白质中含有氨基酸的硫酯的特定于位点的结合。

在这里,我们报告了含有非典型氨基酸(ncAA)的硫酯在重组表达蛋白中的位点特异性结合。具体来说,我们使用正交无义抑制子tRNA /氨酰基-tRNA合成酶(aaRS)对,以高收率和高保真度在细菌中遗传编码了硫酯活化的天冬氨酸(ThioD)。为了证明ThioD的实用性,我们使用天然化学连接以高产率标记带有荧光团的绿色荧光蛋白。
更新日期:2018-01-23
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