An expanded eukaryotic genetic code.
We describe a general and rapid route for the addition of unnatural amino acids to the genetic code of Saccharomyces cerevisiae. Five amino acids have been incorporated into proteins efficiently and with high fidelity in response to the nonsense codon TAG. The side chains of these amino acids contain a keto group, which can be uniquely modified in vitro and in vivo with a wide range of chemical probes and reagents; a heavy atom-containing amino acid for structural studies; and photocrosslinkers for cellular studies of protein interactions. This methodology not only removes the constraints imposed by the genetic code on our ability to manipulate protein structure and function in yeast, it provides a gateway to the systematic expansion of the genetic codes of multicellular eukaryotes.
Chin
Jason W
JW
Department of Chemistry and Skaggs Institute for Chemical Biology, Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Cropp
T Ashton
TA
Anderson
J Christopher
JC
Mukherji
Mridul
M
Zhang
Zhiwen
Z
Schultz
Peter G
PG
D000596:Amino Acids / Q000235:genetics* / Q000378:metabolism; D000926:Anticodon; D001386:Azides / Q000378:metabolism; D018389:Codon, Nonsense; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005815:Genetic Code; D006801:Humans; D008781:Methyltyrosines / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism; D014176:Protein Biosynthesis; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000072105:Superoxide Dismutase-1; D014448:Tyrosine-tRNA Ligase / Q000235:genetics / Q000378:metabolism
C476995:3-(4-acetylphenyl)-2-aminopropanoic acid; D000596:Amino Acids; D000926:Anticodon; D001386:Azides; D018389:Codon, Nonsense; D008781:Methyltyrosines; C000606290:SOD1 protein, human; C002350:O-methyltyrosine; C027162:4-azidophenylalanine; D010649:Phenylalanine; D012343:RNA, Transfer; C053764:benzoylphenylalanine; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D014448:Tyrosine-tRNA Ligase; C049181:4-iodophenylalanine