Genetically Encoded 1,2-Aminothiols Facilitate Rapid and Site-Specific Protein Labeling via a Bio-orthogonal Cyanobenzothiazole Condensation

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• 作者：Duy P. Nguyen ; Thomas Elliott ; Matthew Holt ; Tom W. Muir ; Jason W. Chin
• 刊名：Journal of the American Chemical Society
• 出版年：2011
• 出版时间：August 3, 2011
• 年：2011
• 卷：133
• 期：30
• 页码：11418-11421
• 全文大小：755K
• 年卷期：v.133,no.30(August 3, 2011)
• ISSN：1520-5126

文摘

We report evolved orthogonal pyrrolysyl-tRNA synthetase/tRNA_CUA pairs that direct the efficient, site-specific incorporation of N^蔚-l-thiaprolyl-l-lysine, N^蔚-d-cysteinyl-l-lysine, and N^蔚-l-cysteinyl-l-lysine into recombinant proteins in Escherichia coli. We demonstrate that the unique 1,2-aminothiol introduced by our approach can be efficiently, rapidly, and specifically labeled via a cyanobenzothiazole condensation to quantitatively introduce biophysical probes into proteins. Moreover, we show that, in combination with cysteine labeling, this approach allows the dual labeling of proteins with distinct probes at two distinct, genetically defined sites.