一种代谢烯报告物，用于在哺乳动物细胞中对新合成蛋白质进行时空控制成像。
A metabolic alkene reporter for spatiotemporally controlled imaging of newly synthesized proteins in Mammalian cells.
机构信息
Department of Chemistry, State University of New York at Buffalo, 14260, USA.
出版信息
ACS Chem Biol. 2010 Sep 17;5(9):875-85. doi: 10.1021/cb100193h.
Abstract
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.
摘要
动物细胞中新生蛋白质的非对称空间分布在许多细胞过程中起着核心作用。在这里，我们报告说，一种简单的烯烃标签，即 homoallylglycine（HAG），可以高效地共翻译掺入到重组蛋白以及哺乳动物细胞内的内源性新合成蛋白质中。与光诱导的四唑-烯烃环加成反应（“光点击化学”）结合使用，该烯烃标签还可以作为生物正交化学报告基团，用于体外选择性蛋白质功能化，以及在活哺乳动物细胞中对新合成蛋白质进行时空控制成像。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的蛋白质合成的作用提供一种潜在有用的工具。
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