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.
引用
收藏
页码:875 / 885
页数:11
相关论文
共 52 条
[1]   Chemical Approaches To Perturb, Profile, and Perceive Glycans [J].
Agard, Nicholas J. ;
Bertozzi, Carolyn R. .
ACCOUNTS OF CHEMICAL RESEARCH, 2009, 42 (06) :788-797
[2]   A strain-promoted [3+2] azide-alkyne cycloaddition for covalent modification of blomolecules in living systems [J].
Agard, NJ ;
Prescher, JA ;
Bertozzi, CR .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (46) :15046-15047
[3]   Genetically Encoded Alkenes in Yeast [J].
Ai, Hui-wang ;
Shen, Weijun ;
Brustad, Eric ;
Schultz, Peter G. .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2010, 49 (05) :935-937
[4]   Activity-based protein profiling for the functional annotation of enzymes [J].
Barglow, Katherine T. ;
Cravatt, Benjamin F. .
NATURE METHODS, 2007, 4 (10) :822-827
[5]   Two-color labeling of temporally defined protein populations in mammalian cells [J].
Beatty, Kimberly E. ;
Tirrell, David A. .
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2008, 18 (22) :5995-5999
[6]   Fluorescence visualization of newly synthesized proteins in mammalian cells [J].
Beatty, Kimberly E. ;
Liu, Julie C. ;
Xie, Fang ;
Dieterich, Daniela C. ;
Schuman, Erin M. ;
Wang, Qian ;
Tirrell, David A. .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2006, 45 (44) :7364-7367
[7]   Click Chemistry and Bioorthogonal Reactions: Unprecedented Selectivity in the Labeling of Biological Molecules [J].
Best, Michael D. .
BIOCHEMISTRY, 2009, 48 (28) :6571-6584
[8]   Tetrazine ligation: Fast bioconjugation based on inverse-electron-demand Diels-Alder reactivity [J].
Blackman, Melissa L. ;
Royzen, Maksim ;
Fox, Joseph M. .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2008, 130 (41) :13518-+
[9]   Introducing genetically encoded aldehydes into proteins [J].
Carrico, Isaac S. ;
Carlson, Brian L. ;
Bertozzi, Carolyn R. .
NATURE CHEMICAL BIOLOGY, 2007, 3 (06) :321-322
[10]   Chemical tools for understanding protein lipidation in eukaryotes [J].
Charron, Guillaume ;
Wilson, John ;
Hang, Howard C. .
CURRENT OPINION IN CHEMICAL BIOLOGY, 2009, 13 (04) :382-391