Inducible, selective labeling of proteins via enzymatic oxidation of tyrosine

Jorick J. Bruins, Criss van de Wouw, Jordi F. Keijzer, Bauke Albada*, Floris L. van Delft

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Place of PublicationHertfordshire
PublisherSpringer
Pages357-368
Number of pages12
ISBN (Electronic)9781493995462
ISBN (Print)9781493995455
DOIs
Publication statusPublished - 4 Jun 2019

Publication series

NameMethods in Molecular Biology
Volume2012
ISSN (Print)1064-3745

Keywords

  • Antibody–drug conjugates (ADC)
  • Bioconjugate chemistry
  • Bioorthogonal click
  • Protein functionalization
  • SPOCQ
  • Tyrosine modification

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