Genetically encoded norbornene directs site-specific cellular protein labelling via a rapid bioorthogonal reaction

Kathrin Lang, Lloyd Davis, Jessica Torres-Kolbus, Chungjung Chou, Alexander Deiters, Jason W Chin

Research output: Contribution to journalArticlepeer-review

Abstract

The site-specific incorporation of bioorthogonal groups via genetic code expansion provides a powerful general strategy for site-specifically labelling proteins with any probe. However, the slow reactivity of the bioorthogonal functional groups that can be encoded genetically limits the utility of this strategy. We demonstrate the genetic encoding of a norbornene amino acid using the pyrrolysyl tRNA synthetase/tRNA(CUA) pair in Escherichia coli and mammalian cells. We developed a series of tetrazine-based probes that exhibit 'turn-on' fluorescence on their rapid reaction with norbornenes. We demonstrate that the labelling of an encoded norbornene is specific with respect to the entire soluble E. coli proteome and thousands of times faster than established encodable bioorthogonal reactions. We show explicitly the advantages of this approach over state-of-the-art bioorthogonal reactions for protein labelling in vitro and on mammalian cells, and demonstrate the rapid bioorthogonal site-specific labelling of a protein on the mammalian cell surface.

Original languageEnglish
Pages (from-to)298-304
Number of pages7
JournalNature Chemistry
Volume4
Issue number4
DOIs
Publication statusPublished - Apr 2012

Keywords

  • Amino Acids
  • Amino Acyl-tRNA Synthetases
  • Cell Line
  • Fluorescent Dyes
  • Green Fluorescent Proteins
  • Humans
  • Lysine
  • Norbornanes
  • Proteins
  • Rhodamines

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