Open AccessVisualization of O ‐GlcNAc Glycosylation Stoichiometry and Dynamics Using Resolvable Poly(ethylene glycol) Mass Tags
Author(s) -
Clark Peter M.,
Rexach Jessica E.,
HsiehWilson Linda C.
Publication year - 2013
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch130153
Subject(s) - ethylene glycol , glycosylation , stoichiometry , chemistry , biochemistry , organic chemistry
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.