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.
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Acknowledgments
This work was supported by funding from the University of Pennsylvania and the Searle Scholars Program (10-SSP-214 to EJP). HEG was supported by a summer research fellowship from Eli Lilly.
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Wagner, A.M., Warner, J.B., Garrett, H.E., Walters, C.R., Petersson, E.J. (2015). Transferase-Mediated Labeling of Protein N-Termini with Click Chemistry Handles. In: Kashina, A. (eds) Protein Arginylation. Methods in Molecular Biology, vol 1337. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2935-1_15
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DOI: https://doi.org/10.1007/978-1-4939-2935-1_15
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