Adding amino acids with novel reactivity to the genetic code of Saccharomyces cerevisiae.
Using a novel genetic selection, we have identified a series of mutants of the E. coli tyrosyl-tRNA synthetase that selectively charge an amber suppressor tRNA with p-(propargyloxy)phenylalanine and p-azidophenylalanine in yeast. These evolved tRNA-synthetase pairs can be used to site-specifically label proteins with functional groups orthogonal to normal biological chemistries. As an example, we have shown that proteins containing these amino acids can be efficiently bioconjugated with small organic molecules by a [3 + 2] cycloaddition reaction that is mild enough for the manipulation of biological samples.
D000114:Acetylene / Q000031:analogs & derivatives; D000596:Amino Acids / Q000737:chemistry / Q000235:genetics*; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism; D001386:Azides / Q000737:chemistry; D018389:Codon, Nonsense / Q000235:genetics; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013482:Superoxide Dismutase / Q000235:genetics
D000596:Amino Acids; D001386:Azides; D018389:Codon, Nonsense; D012345:RNA, Transfer, Amino Acid-Specific; D013482:Superoxide Dismutase; D000604:Amino Acyl-tRNA Synthetases; D000114:Acetylene