title: Monitoring protein O-linked β-N-acetylglucosamine status via metabolic labeling and copper-free click chemistry.
abstract: 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.
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA.
authors.forename: Chin Fen
authors.initials: CF
authors.lastname: Teo
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA. Electronic address: lwellsuf@uga.edu.
authors.forename: Lance
authors.initials: L
authors.lastname: Wells
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D057930:Click Chemistry; D003300:Copper / Q000737:chemistry
chemical_list: D003300:Copper; D000117:Acetylglucosamine
keywords: Azido sugars; Cyclooctyne; O-GlcNAc stoichiometry; PEGylation; SPAAC