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 proceedingChapterAcademicpeer-review

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

Fingerprint

Tyrosine
Cycloaddition Reaction
Proteins
Alkynes
Monophenol Monooxygenase
Agaricales
Alkenes
Enzymes
Peptides
Antibodies
benzoquinone

Keywords

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

Cite this

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 (pp. 357-368). (Methods in Molecular Biology; Vol. 2012). Hertfordshire: Springer. https://doi.org/10.1007/978-1-4939-9546-2_18
Bruins, Jorick J. ; van de Wouw, Criss ; Keijzer, Jordi F. ; Albada, Bauke ; van Delft, Floris L. / Inducible, selective labeling of proteins via enzymatic oxidation of tyrosine. Methods in Molecular Biology. Hertfordshire : Springer, 2019. pp. 357-368 (Methods in Molecular Biology).
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Bruins, JJ, van de Wouw, C, Keijzer, JF, Albada, B & van Delft, FL 2019, Inducible, selective labeling of proteins via enzymatic oxidation of tyrosine. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2012, Springer, Hertfordshire, pp. 357-368. https://doi.org/10.1007/978-1-4939-9546-2_18

Inducible, selective labeling of proteins via enzymatic oxidation of tyrosine. / Bruins, Jorick J.; van de Wouw, Criss; Keijzer, Jordi F.; Albada, Bauke; van Delft, Floris L.

Methods in Molecular Biology. Hertfordshire : Springer, 2019. p. 357-368 (Methods in Molecular Biology; Vol. 2012).

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

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Bruins JJ, van de Wouw C, Keijzer JF, Albada B, van Delft FL. Inducible, selective labeling of proteins via enzymatic oxidation of tyrosine. In Methods in Molecular Biology. Hertfordshire: Springer. 2019. p. 357-368. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9546-2_18