Visualization of O‐GlcNAc Glycosylation Stoichiometry and Dynamics Using Resolvable Poly(ethylene glycol) Mass Tags. Issue 4 (16th December 2013) - Clark, Peter M.
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. Curr. Protoc. Chem. Biol . 5:281‐302 © 2013 by John Wiley & Sons, Inc.