Visualization of O-GlcNAc Glycosylation Stoichiometry and Dynamics Using Resolvable Poly(ethylene glycol) Mass Tags
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.
Additional Information
© 2013 John Wiley & Sons, Inc. Published Online: 16 Dec. 2013. We thank P. Qasba (National Cancer Institute at Frederick) for helpful discussions on GalT and for generously providing the Y298L GalT construct. This work was supported by the National Institutes of Health (R01 GM084724 to L.C.H.-W., F31 NS056525 to J.E.R., and 5T32 GM07737 to P.M.C.).Attached Files
Accepted Version - nihms551518.pdf
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Additional details
- PMCID
- PMC3931299
- Eprint ID
- 43347
- Resolver ID
- CaltechAUTHORS:20140113-144333790
- R01 GM084724
- NIH
- F31 NS056525
- NIH Postdoctoral Fellowship
- 5T32 GM07737
- NIH Predoctoral Fellowship
- Created
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2014-01-17Created from EPrint's datestamp field
- Updated
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2021-11-10Created from EPrint's last_modified field