title: Visualization of O-GlcNAc glycosylation stoichiometry and dynamics using resolvable poly(ethylene glycol) mass tags.
abstract: O-linked N-acetylglucosamine (O-GlcNAc) glycosylation is a dynamic protein posttranslational modification with roles in processes such as transcription, cell cycle regulation, and metabolism. Detailed mechanistic studies of O-GlcNAc have been hindered by a lack of methods for measuring O-GlcNAc stoichiometries and the interplay of glycosylation with other posttranslational modifications. We recently developed a method for labeling O-GlcNAc-modified proteins with resolvable poly(ethylene glycol) mass tags. This mass-tagging approach enables the direct measurement of glycosylation stoichiometries and the visualization of distinct O-GlcNAc-modified subpopulations. Here, we describe procedures for labeling O-GlcNAc glycoproteins in cell lysates with mass tags.
authors.affiliation: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Clark
authors.forename: Jessica E
authors.initials: JE
authors.lastname: Rexach
authors.forename: Linda C
authors.initials: LC
authors.lastname: Hsieh-Wilson
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D000818:Animals; D001386:Azides / Q000737:chemistry; D002240:Carbohydrate Sequence; D061565:Cycloaddition Reaction / Q000379:methods; D005700:Galactosyltransferases / Q000737:chemistry / Q000235:genetics; D006031:Glycosylation / Q000187:drug effects*; D006801:Humans; D008969:Molecular Sequence Data; D010091:Oximes / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011499:Protein Processing, Post-Translational / Q000235:genetics; D011506:Proteins / Q000737:chemistry
chemical_list: D001386:Azides; D010091:Oximes; D011506:Proteins; D011092:Polyethylene Glycols; D005700:Galactosyltransferases; D000117:Acetylglucosamine
keywords: O-linked N-acetylglucosamine; chemoenzymatic labeling; glycosylation; poly(ethylene glycol); posttranslational modifications; protein subpopulations