Two Rapid Catalyst-Free Click Reactions for In Vivo Protein Labeling of Genetically Encoded Strained Alkene/Alkyne Functionalities

Detailed kinetic analyses of inverse electron-demand Diels–Alder 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/alky...

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Bibliographic Details
Published in:Bioconjugate chemistry 2014-09, Vol.25 (9), p.1730-1738
Main Authors: Kurra, Yadagiri, Odoi, Keturah A, Lee, Yan-Jiun, Yang, Yanyan, Lu, Tongxiang, Wheeler, Steven E, Torres-Kolbus, Jessica, Deiters, Alexander, Liu, Wenshe R
Format: Article
Language:English
Subjects:
Alkenes - chemistry
Alkynes - chemistry
Catalysis
Click Chemistry
Cyclooctanes - chemistry
Escherichia coli Proteins - chemistry
Kinetics
Models, Molecular
Protein Conformation
Proteins
Proteins - chemistry
Staining and Labeling - methods
Tyrosine - chemistry
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Summary:Detailed kinetic analyses of inverse electron-demand Diels–Alder 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-tRNACUA 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.
ISSN:1043-1802
1520-4812
DOI:10.1021/bc500361d