Genetic Incorporation of Histidine Derivatives Using an Engineered Pyrrolysyl-tRNA Synthetase
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- 作者：Han Xiao ; Francis B. Peters ; Peng-Yu Yang ; Sean Reed ; Johnathan R. Chittuluru ; Peter G. Schultz
- 刊名：ACS Chemical Biology
- 出版年：2014
- 出版时间：May 16, 2014
- 年：2014
- 卷：9
- 期：5
- 页码：1092-1096
- 全文大小：259K
- 年卷期：v.9,no.5(May 16, 2014)
- ISSN：1554-8937
文摘
A polyspecific amber suppressor aminoacyl-tRNA synthetase/tRNA pair was evolved that genetically encodes a series of histidine analogues in both Escherichia coli and mammalian cells. In combination with tRNACUAPyl, a pyrrolysyl-tRNA synthetase mutant was able to site-specifically incorporate 3-methyl-histidine, 3-pyridyl-alanine, 2-furyl-alanine, and 3-(2-thienyl)-alanine into proteins in response to an amber codon. Substitution of His66 in the blue fluorescent protein (BFP) with these histidine analogues created mutant proteins with distinct spectral properties. This work further expands the structural and chemical diversity of unnatural amino acids (UAAs) that can be genetically encoded in prokaryotic and eukaryotic organisms and affords new probes of protein structure and function.