Monitoring protein O-linked β-N-acetylglucosamine status via metabolic labeling and copper-free click chemistry

Anal Biochem. 2014 Nov 1;464:70-2. doi: 10.1016/j.ab.2014.06.010. Epub 2014 Jul 1.


O-Linked β-N-acetylglucosamine (O-GlcNAc) modification found on the serine and threonine residues of intracellular proteins is an inducible post-translational modification that regulates numerous biological processes. In combination with other cell biological and biochemical approaches, a robust and streamlined strategy for detecting the number and stoichiometry of O-GlcNAc modification can provide valuable insights for decoding the functions of O-GlcNAc at the molecular level. Here, we report an optimized workflow for evaluating the O-GlcNAc status of proteins using a combination of metabolic labeling and click chemistry-based mass tagging. This method is strategically complementary to the chemoenzymatic-based mass-tagging method.

Keywords: Azido sugars; Cyclooctyne; O-GlcNAc stoichiometry; PEGylation; SPAAC.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetylglucosamine / metabolism*
  • Click Chemistry*
  • Copper / chemistry


  • Copper
  • Acetylglucosamine