In vivo incorporation of an alkyne into proteins in Escherichia coli.
Using a genetic selection we identified mutants of the M. janaschii tyrosyl-tRNA synthetase that selectively charge an amber suppressor tRNA with para-propargyloxyphenylalanine in Escherichia coli. These evolved tRNA-synthetase pairs were used to site-specifically incorporate an alkynyl group into a protein, which was subsequently conjugated with fluorescent dyes by a [3+2]-cycloaddition reaction under mild reaction conditions.
Deiters
Alexander
A
Department of Chemistry, Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Schultz
Peter G
PG
D000480:Alkynes / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D003500:Cyclization; D019020:Directed Molecular Evolution; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry; D015394:Molecular Structure; D009154:Mutation; D009211:Myoglobin / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000378:metabolism; D015202:Protein Engineering; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D014448:Tyrosine-tRNA Ligase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014907:Whales
D000480:Alkynes; D005456:Fluorescent Dyes; D009211:Myoglobin; D011506:Proteins; D010649:Phenylalanine; D012343:RNA, Transfer; D014448:Tyrosine-tRNA Ligase