A genetically encoded fluorescent probe in mammalian cells

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 resp...

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Bibliographic Details
Published in:Journal of the American Chemical Society Vol. 135; no. 34; p. 12540
Main Authors: Chatterjee, Abhishek, Guo, Jiantao, Lee, Hyun Soo, Schultz, Peter G
Format: Journal Article
Language:English
Published: United States 28.08.2013
Subjects:
Amino Acyl-tRNA Synthetases - chemistry
Amino Acyl-tRNA Synthetases - genetics
Amino Acyl-tRNA Synthetases - metabolism
Animals
beta-Alanine - analogs & derivatives
beta-Alanine - chemistry
beta-Alanine - genetics
beta-Alanine - metabolism
CHO Cells
Cricetulus
Fluorescent Dyes - chemistry
Fluorescent Dyes - metabolism
HEK293 Cells
HeLa Cells
Humans
RNA, Transfer - chemistry
RNA, Transfer - genetics
ISSN:1520-5126, 1520-5126
Online Access:Get more information
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Summary: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.
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ISSN:1520-5126
1520-5126
DOI:10.1021/ja4059553