Two rapid catalyst-free click reactions for in vivo protein labeling of genetically encoded strained alkene/alkyne functionalities.
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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.
author list (cited authors)
Kurra, Y., Odoi, K. A., Lee, Y., Yang, Y., Lu, T., Wheeler, S. E., ... Liu, W. R.
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