A metabolic alkene reporter for spatiotemporally controlled imaging of newly synthesized proteins in Mammalian cells.

A metabolic alkene reporter for spatiotemporally controlled imaging of newly synthesized proteins in Mammalian cells.
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DOI:
10.1021/cb100193h
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发表时间:
2010-09-17
影响因子:
4
通讯作者:
Lin, Qing
Lin, Qing
中科院分区:
生物学2区
文献类型:
--
作者:
Song, Wenjiao;Wang, Yizhong;Yu, Zhipeng;Vera, Claudia I. Rivera;Qu, Jun;Lin, Qing

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动物细胞中新合成蛋白质的非对称空间分布在许多细胞过程中发挥着核心作用。在这里,我们报告了一个简单的烯烃标签,高烯丙基甘氨酸(HAG),被高效地共翻译整合到重组蛋白以及哺乳动物细胞中的内源性新合成蛋白中。与光诱导的四唑-烯烃环加成反应(“光点击化学”)相结合,该烯烃标签进一步充当生物正交化学报告分子,用于体外选择性蛋白质功能化和活哺乳动物细胞中新合成蛋白质的时空控制成像。这种两步代谢烯烃标记-光控化学功能化方法可能为研究哺乳动物细胞中时空调节蛋白质合成的作用提供潜在有用的工具。
The non-symmetrical 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–photo-controlled chemical functionalization approach may offer a potentially useful tool to study the role of the spatiotemporally regulated protein synthesis in mammalian cells.
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