Site-Specific Glycoconjugation of Protein via Bioorthogonal Tetrazine Cycloaddition with a Genetically Encoded trans-Cyclooctene or Bicyclononyne

Bioconjug Chem. 2015 May 20;26(5):802-6. doi: 10.1021/acs.bioconjchem.5b00101. Epub 2015 Apr 24.

Abstract

Efficient access to proteins modified site-specifically with glycans is important in glycobiology and for therapeutic applications. Herein, we report a biocompatible protein glycoconjugation by inverse demand Diels-Alder reaction between tetrazine and trans-cyclooctene. Tetrazine functionalized glycans were obtained in one step by CuAAC (Cu-catalyzed alkyne azide cycloaddition) between glycosyl azide and an alkyne-tetrazine adduct. Site-specific glycoconjugation was performed chemoselectively on a target protein in which a trans-cyclooctene derivatized lysine was genetically encoded. Glycoconjugation proceeded to completion on purified protein and was shown to be selective for the target protein in E. coli.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkynes
  • Azides / chemistry
  • Binding Sites
  • Catalysis
  • Copper / chemistry
  • Cycloaddition Reaction*
  • Cyclooctanes / chemistry*
  • Cycloparaffins / chemistry*
  • Escherichia coli / genetics
  • Glycoproteins / chemistry*
  • Glycoproteins / genetics*
  • Models, Molecular
  • Protein Conformation
  • Substrate Specificity

Substances

  • Alkynes
  • Azides
  • Cyclooctanes
  • Cycloparaffins
  • Glycoproteins
  • Copper