A Metabolic Alkene Reporter for Spatiotemporally Controlled Imaging of Newly Synthesized Proteins in Mammalian Cells

被引：76

作者：

Song, Wenjiao ^{[1
]}

Wang, Yizhong ^{[1
]}

Yu, Zhipeng ^{[1
]}

Vera, Claudia I. Rivera ^{[1
]}

Qu, Jun ^{[2
]}

Lin, Qing ^{[1
]}

机构：

[1] SUNY Buffalo, Dept Chem, Buffalo, NY 14260 USA

[2] SUNY Buffalo, Dept Pharmaceut Sci, Buffalo, NY 14260 USA

来源：

ACS CHEMICAL BIOLOGY | 2010年 / 5卷 / 09期

基金：

美国国家卫生研究院;

关键词：

TRANSFER-RNA SYNTHETASE; IN-VIVO; PHOTOCLICK CHEMISTRY; ESCHERICHIA-COLI; CYCLOADDITION; LIGATION; FUNCTIONALIZATION; VISUALIZATION; SELECTIVITY; REACTIVITY;

D O I：

10.1021/cb100193h

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：

The nonsymmetrical spatial distribution of newly synthesized proteins in animal cells plays a central role in many cellular processes. Here, we report that a simple alkene tag, homoallylglycine (HAG), was co-translationally incorporated into a recombinant protein as well as endogenous, newly synthesized proteins in mammalian cells with high efficiency. In conjunction with a photoinduced tetrazole-alkene cycloaddition reaction ("photoclick chemistry"), this alkene tag further served as a bioorthogonal chemical reporter both for the selective protein functionalization in vitro and for a spatiotemporally controlled imaging of the newly synthesized proteins in live mammalian cells. This two-step metabolic alkene tagging-photocontrolled chemical functionalization approach may offer a potentially useful tool to study the role of spatiotemporally regulated protein synthesis in mammalian cells.

引用

页码：875 / 885

页数：11