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Curr Protoc Chem Biol. 2013;5(4):281-302. doi: 10.1002/9780470559277.ch130153.

Visualization of O-GlcNAc glycosylation stoichiometry and dynamics using resolvable poly(ethylene glycol) mass tags.

Author information

1
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California.

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.

KEYWORDS:

O-linked N-acetylglucosamine; chemoenzymatic labeling; glycosylation; poly(ethylene glycol); posttranslational modifications; protein subpopulations

PMID:
24391098
PMCID:
PMC3931299
DOI:
10.1002/9780470559277.ch130153
[Indexed for MEDLINE]
Free PMC Article

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