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 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.
Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Abhishek
A
Chatterjee
Jiantao
J
Guo
Hyun Soo
HS
Lee
Peter G
PG
Schultz
D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics*; D015091:beta-Alanine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
C546053:3-(6-acetylnaphthalen-2-ylamino)-2-aminopropanoic acid; D005456:Fluorescent Dyes; D015091:beta-Alanine; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases