MassIVE MSV000087290

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Novel antibodies for the simple and efficient enrichment of native O-GlcNAc modified peptides

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Description

Antibodies against post-translational modifications (PTMs) such as lysine acetylation, ubiquitnation, or phospho-tyrosine have enabled significant advances in our understanding the fundamental roles of PTMs in biology. However, a number of PTM spaces remain unexplored due to the lack of robust enrichment reagents. The addition of N-acetylglucosamine to serine and threonine residues (O-GlcNAc) is a PTM implicated in numerous biological processes and disease states, and has limited techniques for its study. Here, we evaluate a new mixture of anti-O-GlcNAc monoclonal antibodies for the immunoprecipitation of native O-GlcNAcylated peptides from cells and tissues. The anti-O-GlcNAc antibodies display good sensitivity and excellent specificity toward O-GlcNAc, and not O-GalNAc or GlcNAc in extended glycans. Applying these antibodies to tissue samples, we achieved an in-depth characterization of O-GlcNAcylation from mouse synaptosomes, identifying over 1,300 unique O-GlcNAc-modified peptides and over 1,000 sites, using a fraction of sample preparation and instrument time of other landmark investigations. This rapid and robust method greatly simplifies the analysis of O-GlcNAc signaling and will help to elucidate the role of this challenging PTM in health and disease. [doi:10.25345/C5TV5G] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: O-GlcNAc ; Antibodies ; ETD ; HCD-pd-ETD ; embryonic stem cells ; synaptosomes

Contact

Principal Investigators:
(in alphabetical order) 		Samuel Myers, La Jolla Institute for Immunology, United States
Submitting User: sammyers
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