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  • Title: Inducible, Selective Labeling of Proteins via Enzymatic Oxidation of Tyrosine.
    Author: Bruins JJ, van de Wouw C, Keijzer JF, Albada B, van Delft FL.
    Journal: Methods Mol Biol; 2019; 2012():357-368. PubMed ID: 31161517.
    Abstract:
    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.
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