Site-Specific Incorporation of a Thioester Containing Amino Acid into Proteins

被引：23

作者：

Xuan, Weimin ^{[1
,2
]}

Collins, Daniel ^{[3
]}

Koh, Minseob ^{[1
,2
]}

Shao, Sida ^{[1
,2
]}

Yao, Anzhi ^{[1
,2
]}

Xiao, Han ^{[1
,2
]}

Garner, Philip ^{[3
]}

Schultz, Peter G. ^{[1
,2
]}

机构：

[1] Scripps Res Inst, Dept Chem, 10550 N Torrey Pines Rd, La Jolla, CA 92037 USA

[2] Scripps Res Inst, Skaggs Inst Chem Biol, 10550 N Torrey Pines Rd, La Jolla, CA 92037 USA

[3] Washington State Univ, Dept Chem, Pullman, WA 99164 USA

来源：

ACS CHEMICAL BIOLOGY | 2018年 / 13卷 / 03期

基金：

美国国家卫生研究院; 美国国家科学基金会;

关键词：

NATIVE CHEMICAL LIGATION; TRANSFER-RNA SYNTHETASE; GENETIC-CODE; IN-VIVO; UBIQUITINATION; CONJUGATION; MUTAGENESIS; EXPANSION;

D O I：

10.1021/acschembio.7b00998

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：578 / 581

页数：4

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