一种用于哺乳动物细胞的基因编码荧光探针。
A genetically encoded fluorescent probe in mammalian cells.
作者信息
Chatterjee Abhishek, Guo Jiantao, Lee Hyun Soo, Schultz Peter G
机构信息
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
出版信息
J Am Chem Soc. 2013 Aug 28;135(34):12540-3. doi: 10.1021/ja4059553. Epub 2013 Aug 14.
Abstract
Fluorescent reporters are useful in vitro and in vivo probes of protein structure, function, and localization. Here we report that the fluorescent amino acid, 3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid (Anap), can be site-specifically incorporated into proteins in mammalian cells in response to the TAG codon with high efficiency using an orthogonal amber suppressor tRNA/aminoacyl-tRNA synthetase (aaRS) pair. We further demonstrate that Anap can be used to image the subcellular localization of proteins in live mammalian cells. The small size of Anap, its environment-sensitive fluorescence, and the ability to introduce Anap at specific sites in the proteome by simple mutagenesis make it a unique and valuable tool in eukaryotic cell biology.
摘要
荧光报告分子是用于蛋白质结构、功能及定位研究的有用的体外和体内探针。在此我们报告，荧光氨基酸3-(6-乙酰萘-2-基氨基)-2-氨基丙酸（Anap），可利用正交琥珀抑制tRNA/氨酰-tRNA合成酶（aaRS）对，高效地响应TAG密码子在哺乳动物细胞中位点特异性地掺入蛋白质中。我们进一步证明，Anap可用于对活的哺乳动物细胞中蛋白质的亚细胞定位进行成像。Anap体积小、具有环境敏感型荧光，并且能够通过简单诱变在蛋白质组的特定位点引入Anap，这使其成为真核细胞生物学中一种独特且有价值的工具。
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