共 52 条
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
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.
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页码:875 / 885
页数:11
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