Visualization of O‐GlcNAc Glycosylation Stoichiometry and Dynamics Using Resolvable Poly(ethylene glycol) Mass Tags. Issue 4 (16th December 2013)

• Record Type: Journal Article
• Authors: Clark, Peter M.; Rexach, Jessica E.; Hsieh‐Wilson, Linda C.
• Editors: Mahal, Lara; Romesberg, Floyd; Shah, Kavita; Shamu, Caroline; Strano, Michael S.; Thomas, Craig
• Is Part Of: Current protocols in chemical biology. Volume 5:Issue 4(2013)
• Page Start: 281
• Page End: 302
• Publication Date: 2013-12-16
• Subjects: O‐linked N‐acetylglucosamine -- glycosylation -- posttranslational modifications -- chemoenzymatic labeling -- poly(ethylene glycol) -- protein subpopulations; Biochemistry -- Laboratory manuals; Chemical Phenomena; Biochemistry; Periodicals; Laboratory manuals; 572.078
• DOI: 10.1002/9780470559277.ch130153
• Languages: English
• ISSNs: 2160-4762

Abstract

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.