Monitoring protein O-linked β-N-acetylglucosamine status via metabolic labeling and copper-free click chemistry.
Analytical Biochemistry(2014)
摘要
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.
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关键词
O-GlcNAc stoichiometry,Azido sugars,SPAAC,Cyclooctyne,PEGylation
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