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Visualization of O-GlcNAc Glycosylation Stoichiometry and Dynamics using Resolvable Poly(ethylene glycol) Mass Tags
Journal article   Open access

Visualization of O-GlcNAc Glycosylation Stoichiometry and Dynamics using Resolvable Poly(ethylene glycol) Mass Tags

Peter M. Clark, Jessica E. Rexach and Linda C. Hsieh-Wilson
Current protocols in chemical biology, v 5(4), pp 281-302
01 Dec 2013
PMID: 24391098
url
https://europepmc.org/articles/pmc3931299View
Accepted (AM)Open Access (License Unspecified) Open

Abstract

chemoenzymatic labeling glycosylation O-linked N-acetylglucosamine poly(ethylene glycol) posttranslational modifications protein subpopulations
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 protocols for labeling O -GlcNAc glycoproteins in cell lysates with mass tags.

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