Site-specific labeling of proteins and peptides with trans-cyclooctene containing handles capable of tetrazine ligation

James W. Wollack, Benjamin J. Monson, Jonathan K. Dozier, Joseph J. Dalluge, Kristina Poss, Scott A. Hilderbrand, Mark D. Distefano

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

There is a growing library of functionalized non-natural substrates for the enzyme protein farnesyltransferase (PFTase). PFTase covalently attaches these functionalized non-natural substrates to proteins ending in the sequence CAAX, where C is a cysteine that becomes alkylated, A represents an aliphatic amino acid, and X is Ser, Met, Ala, or Gln. Reported substrates include a variety of functionalities that allow modified proteins to undergo subsequent bioconjugation reactions. To date the most common strategy used in this approach has been copper catalyzed azide-alkyne cycloaddition (CuAAC). While being fast and bioorthogonal CuAAC has limited use in live cell experiments due to copper's toxicity.1 Here, we report the synthesis of trans-cyclooctene geranyl diphosphate. This substrate can be synthesized from geraniol in six steps and be enzymatically transferred to peptides and proteins that end in a CAAX sequence. Proteins and peptides site-specially modified with trans-cyclooctene geranyl diphosphate were subsequently targeted for further modification via tetrazine ligation. As tetrazine ligation is bioorthogonal, fast, and is contingent on ring strain rather than the addition of a copper catalyst, this labeling strategy should prove useful for labeling proteins where the presence of copper may hinder solubility or biological reactivity.

Original languageEnglish (US)
Pages (from-to)140-147
Number of pages8
JournalChemical Biology and Drug Design
Volume84
Issue number2
DOIs
StatePublished - Aug 2014

Keywords

  • bioorthogonal
  • inverse-electron-demand Diels-Alder
  • protein farnesyltransferase
  • protein prenylation
  • tetrazine ligation
  • trans-cyclooctene

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