1. Transferase-Mediated Labeling of Protein N-Termini with Click Chemistry Handles
- Author
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E. James Petersson, Anne Wagner, Haviva E. Garrett, Christopher R. Walters, and John B. Warner
- Subjects
chemistry.chemical_classification ,fungi ,food and beverages ,Alkyne ,A protein ,Biology ,Amino acid ,chemistry.chemical_compound ,chemistry ,Biotin ,Biochemistry ,Click chemistry ,Transferase ,Azide - 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 method can be used to directly detect AaT substrates or in a two-step protocol to detect substrates of the mammalian ATE1 transferase.
- Published
- 2023
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