Copper-Free Click Enabled Triazabutadiene for Bioorthogonal Protein Functionalization

Bioconjug Chem. 2021 Feb 17;32(2):254-258. doi: 10.1021/acs.bioconjchem.0c00677. Epub 2021 Jan 25.

Abstract

Aryl diazonium ions have long been used in bioconjugation due to their reactivity toward electron-rich aryl residues, such as tyrosine. However, their utility in biological systems has been restricted due to the requirement of harsh conditions for their generation in situ, as well as limited hydrolytic stability. Previous work describing a scaffold known as triazabutadiene (TBD) has shown the ability to protect aryl diazonium ions allowing for increased synthetic utility, as well as triggered release under biologically relevant conditions. Herein, we describe the synthesis and application of a novel TBD, capable of installation of a cyclooctyne on protein surfaces for later use of copper-free click reactions involving functional azides. The probe shows efficient protein labeling across a wide pH range that can be accomplished in a convenient and timely manner. Orthogonality of the cyclooctyne modification was showcased by labeling a model protein in the presence of hen egg proteins, using an azide-containing fluorophore. We further confirmed that the azobenzene modification can be cleaved using sodium dithionite treatment.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Butadienes / chemistry*
  • Click Chemistry*
  • Copper / chemistry*
  • Electrophoresis, Polyacrylamide Gel
  • Proteins / chemistry*

Substances

  • Butadienes
  • Proteins
  • Copper