Two rapid catalyst-free click reactions for in vivo protein labeling of genetically encoded strained alkene/alkyne functionalities. Academic Article uri icon


  • Detailed kinetic analyses of inverse electron-demand DielsAlder cycloaddition and nitrilimine-alkene/alkyne 1,3-diploar cycloaddition reactions were conducted and the reactions were applied for rapid protein bioconjugation. When reacted with a tetrazine or a diaryl nitrilimine, strained alkene/alkyne entities including norbornene, trans-cyclooctene, and cyclooctyne displayed rapid kinetics. To apply these click reactions for site-specific protein labeling, five tyrosine derivatives that contain a norbornene, trans-cyclooctene, or cyclooctyne entity were genetically encoded into proteins in Escherichia coli using an engineered pyrrolysyl-tRNA synthetase-tRNA(CUA)(Pyl) pair. Proteins bearing these noncanonical amino acids were successively labeled with a fluorescein tetrazine dye and a diaryl nitrilimine both in vitro and in living cells.

published proceedings

  • Bioconjug Chem

altmetric score

  • 0.75

author list (cited authors)

  • Kurra, Y., Odoi, K. A., Lee, Y., Yang, Y., Lu, T., Wheeler, S. E., ... Liu, W. R.

citation count

  • 55

complete list of authors

  • Kurra, Yadagiri||Odoi, Keturah A||Lee, Yan-Jiun||Yang, Yanyan||Lu, Tongxiang||Wheeler, Steven E||Torres-Kolbus, Jessica||Deiters, Alexander||Liu, Wenshe R

publication date

  • September 2014