Transferase-Mediated Labeling of Protein N-Termini with Click Chemistry Handles

• DOI: 10.1007/978-1-0716-2942-0_21
• Publication Date: 2023-04-03T12:31:25Z
• Authors: Anne M. Wagner; John B. Warner; Haviva E. Garrett; Christopher R. Wa...; E. James Petersson
• Keywords: Transferase; Alkyne; Bioorthogonal Chemistry

The E. coli aminoacyl transferase (AaT) can be used to transfer a variety of unnatural amino acids, including those with azide or alkyne groups, to the α-amine of a protein with an N-terminal Lys or Arg. Subsequent functionalization through either copper-catalyzed or strain-promoted click reactions can be used to label the protein with fluorophores or biotin. This method can be used to directly detect AaT substrates or in a two-step protocol to detect substrates of the mammalian ATE1 transferase.