Ligation of Expressed Protein alpha-Hydrazides via Genetic Incorporation   of an alpha-Hydroxy Acid

doi:10.1021/cb300020s

CORC > 北京大学 > 化学与分子工程学院

	Ligation of Expressed Protein alpha-Hydrazides via Genetic Incorporation of an alpha-Hydroxy Acid
	Li, Yi-Ming ; Yang, Mai-Yun ; Huang, Yi-Chao ; Li, Yi-Tong ; Chen, Peng R. ; Liu, Lei
刊名	acs chemical biology
	2012
关键词	UNNATURAL AMINO-ACIDS ACTIVE-ESTER-CONDENSATION TRANSFER-RNA SYNTHETASE CHEMICAL-SYNTHESIS PERIPLASMIC PROTEIN SIGNALING PROTEIN ENTERIC BACTERIA MAMMALIAN-CELLS CODE HDEA
DOI	10.1021/cb300020s
英文摘要	Expressed protein ligation bridges the gap between synthetic peptides and recombinant proteins and thereby significantly increases the size and complexity of chemically synthesized proteins. Although the intein-based expressed protein ligation method has been extensively used in this regard, the development of new expressed protein ligation methods may improve the flexibility and power of protein semisynthesis. In this study a new alternative version of expressed protein ligation is developed by combining the recently developed technologies of hydrazide-based peptide ligation and genetic code expansion. Compared to the previous intein-based expressed protein ligation method, the new method does not require the use of protein splicing technology and generates recombinant protein alpha-hydrazides as ligation intermediates that are more chemically stable than protein alpha-thioesters. Furthermore, the use of an evolved mutant pyrrolysyl-tRNA synthetase (PylRS), ACPK-RS, from M. barkeri shows an improved performance for the expression of recombinant protein backbone oxoesters. By using HdeA as a model protein we demonstrate that the hydrazide-based method can be used to synthesize proteins with correctly folded structures and full biological activity. Because the PylRS-tRNA(CUA)(pyl) system is compatible with both prokaryotic and eukaryotic cells, the strategy presented here may be readily expanded to manipulate proteins produced in mammalian cells. The new hydrazide-based method may also supplement the intein-based expressed protein ligation method by allowing for a more flexible selection of ligation site.; Biochemistry & Molecular Biology; SCI(E); 0; ARTICLE; 6; 1015-1022; 7
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/232854]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	Li, Yi-Ming,Yang, Mai-Yun,Huang, Yi-Chao,et al. Ligation of Expressed Protein alpha-Hydrazides via Genetic Incorporation of an alpha-Hydroxy Acid[J]. acs chemical biology,2012.
APA	Li, Yi-Ming,Yang, Mai-Yun,Huang, Yi-Chao,Li, Yi-Tong,Chen, Peng R.,&Liu, Lei.(2012).Ligation of Expressed Protein alpha-Hydrazides via Genetic Incorporation of an alpha-Hydroxy Acid.acs chemical biology.
MLA	Li, Yi-Ming,et al."Ligation of Expressed Protein alpha-Hydrazides via Genetic Incorporation of an alpha-Hydroxy Acid".acs chemical biology (2012).