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Mass Spectrometry-Based Quantitative O-GlcNAcomic Analysis

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Quantitative Proteomics by Mass Spectrometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1410))

Abstract

The dynamic co- and post-translational modification (PTM) of proteins, O-linked β-d-N-acetylglucosamine modification (O-GlcNAcylation) of serine/threonine residues is critical in many cellular processes, contributing to multiple physiological and pathological events. The term “O-GlcNAcome” refers to not only the complete set of proteins that undergo O-GlcNAcylation but also the O-GlcNAc status at individual residues, as well as the dynamics of O-GlcNAcylation in response to various stimuli. O-GlcNAcomic analyses have been a challenge for many years. In this chapter, we describe a recently developed approach for the identification and quantification of O-GlcNAc proteins/peptides from complex samples.

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Acknowledgments

Original research in this work was supported by NIH P01HL107153, R01DK61671, and NIH N01-HV-00240 (to G.W.H.). We appreciate Dr. Zihao Wang for his significant contribution for the initial development of this protocol and for his critical reading of the manuscript. Helpful discussion from the Hart laboratory is acknowledged. We also thank Drs. Feng Yang and Richard Smith and their coworkers at the Pacific Northwest National Laboratory (Richland, WA) for valuable comments of the enrichment protocol.

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Authors and Affiliations

  1. Department of Biological Chemistry, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, MD, 21205-2185, USA

    Junfeng Ma & Gerald W. Hart Ph.D.

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  1. Junfeng Ma
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  2. Gerald W. Hart Ph.D.
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Correspondence to Gerald W. Hart Ph.D. .

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Editors and Affiliations

  1. NIDDK, National Institutes of Health, BETHESDA, Maryland, USA

    Salvatore Sechi

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Ma, J., Hart, G.W. (2016). Mass Spectrometry-Based Quantitative O-GlcNAcomic Analysis. In: Sechi, S. (eds) Quantitative Proteomics by Mass Spectrometry. Methods in Molecular Biology, vol 1410. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3524-6_6

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  • DOI: https://doi.org/10.1007/978-1-4939-3524-6_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3522-2

  • Online ISBN: 978-1-4939-3524-6

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