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A genetically encoded fluorescent probe in mammalian cells.

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.

SUBMITTER: Chatterjee A

PROVIDER: S-EPMC3783214 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

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A genetically encoded fluorescent probe in mammalian cells.

Chatterjee Abhishek A Guo Jiantao J Lee Hyun Soo HS Schultz Peter G PG

Journal of the American Chemical Society 20130814 34

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 locali ...[more]

PMID: 23924161

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