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. 2023:2620:157-175.
doi: 10.1007/978-1-0716-2942-0_21.

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

Anne M Wagner  1 , John B Warner  1 , Haviva E Garrett  1 , Christopher R Walters  1 , E James Petersson  2
Affiliations

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

Anne M Wagner et al. Methods Mol Biol. 2023.

Abstract

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 can be used to directly detect AaT substrates or in a two-step protocol to detect substrates of the mammalian ATE1 transferase.

Keywords: Aminoacyl transferase; Chemoenzymatic reaction; Click chemistry; N-end rule; N-terminus; Protein labeling; Protein modification.

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