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Cited by 261

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Enrichment and Site Mapping of O-Linked N-Acetylglucosamine by a Combination of Chemical/Enzymatic Tagging, Photochemical Cleavage, and Electron Transfer Dissociation Mass Spectrometry

Open access
Wang, Zihao, Udeshi, Namrata D., O'Malley, Meaghan, Shabanowitz, Jeffrey, Hunt, Donald F., Hart, Gerald W.·2010·Molecular & Cellular Proteomics

Abstract

Numerous cellular processes are regulated by the reversible addition of either phosphate or O-linked beta-N-acetylglucosamine (O-GlcNAc) to nuclear and cytoplasmic proteins. Although sensitive methods exist for the enrichment and identification of protein phosphorylation sites, those for the enrichment of O-GlcNAc-containing peptides are lacking. Reported here is highly efficient methodology for the enrichment and characterization of O-GlcNAc sites from complex samples. In this method, O-GlcNAc-modified peptides are tagged with a novel biotinylation reagent, enriched by affinity chromatography, released from the solid support by photochemical cleavage, and analyzed by electron transfer dissociation mass spectrometry. Using this strategy, eight O-GlcNAc sites were mapped from a tau-enriched sample from rat brain. Sites of GlcNAcylation were characterized on important neuronal proteins such as tau, synucleins, and methyl CpG-binding protein 2.

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Metadata

Authors
Wang, Z., Udeshi, N. D., O'Malley, M., Shabanowitz, J., Hunt, D. F., & Hart, G. W.
Venue
Molecular & Cellular Proteomics
Publisher
Elsevier BV
Published
2010-01
Volume
9
Issue
1
Pages
153-160
ISSN
1535-9476 (ISSN-L)
OpenAlex
W2154522012
Cited by
261
References
6
FWCI
6.96
Type
journal article
Language
en
Open access
hybrid

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CoChat. (2026). Citation check: Enrichment and Site Mapping of O-Linked N-Acetylglucosamine by a Combination of Chemical/Enzymatic Tagging, Photochemical Cleavage, and Electron Transfer Dissociation Mass Spectrometry. https://app.cochat.ai/papers/10.1074/mcp.m900268-mcp200
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Wang, Z., Udeshi, N. D., O'Malley, M., Shabanowitz, J., Hunt, D. F., & Hart, G. W. (2010). Enrichment and Site Mapping of O-Linked N-Acetylglucosamine by a Combination of Chemical/Enzymatic Tagging, Photochemical Cleavage, and Electron Transfer Dissociation Mass Spectrometry. Molecular & Cellular Proteomics. https://doi.org/10.1074/mcp.m900268-mcp200

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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

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