Proteins can be labeled site-specifically and in inducible fashion by exposing a small peptide tag (G4Y) on any of its termini and activating the newly exposed tyrosine residue with the enzyme mushroom tyrosinase. The enzyme generates a quinone by oxidizing the tyrosine, which in turn can perform strain-promoted oxidation-controlled ortho-quinone cycloaddition (SPOCQ) with strained alkynes and alkenes, generating a stable conjugation product. Here, we describe a protocol to perform SPOCQ reaction on proteins, along with notes to optimize yield and reaction rates. Conjugation efficiencies of over 95% to antibodies have been reported using this protocol.
CITATION STYLE
Bruins, J. J., van de Wouw, C., Keijzer, J. F., Albada, B., & van Delft, F. L. (2019). Inducible, selective labeling of proteins via enzymatic oxidation of tyrosine. In Methods in Molecular Biology (Vol. 2012, pp. 357–368). Humana Press Inc. https://doi.org/10.1007/978-1-4939-9546-2_18
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