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Stable Isotope Labeling of N-Glycosylated Peptides by Enzymatic Deglycosylation for Mass Spectrometry-Based Glycoproteomics

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Mass Spectrometry of Glycoproteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 951))

Abstract

Protein glycosylation is one of the most common and crucial post-translational modifications that regulates many biological processes. Because abnormal glycosylation is also associated with various pathologies, including cancer, and inflammatory and degenerative diseases, technology for comprehensive analysis of glycoproteins, or glycoproteomics, is important not only for biological studies but also for biomedical and clinical research, including the discovery of biomarkers for disease diagnosis, prognosis, and therapeutic response to drugs. Here, we describe a protocol for peptide-N-glycanase-mediated 18O labeling of N-glycosylated peptides, termed “isotope-coded glycosylation site-specific tagging.” Coupled with advanced mass spectrometry-based proteomics technology, this method facilitates the identification of hundreds to thousands of N-glycoproteins, coupled with their sites of glycosylation, from a complex biological mixture.

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Author information

Authors and Affiliations

  1. Department of Chemistry, Tokyo Metropolitan University, Tokyo, Japan

    Hiroyuki Kaji

  2. Department of Chemistry, Tokyo Metropolitan University, and CREST, Japan Science and Technology Agency (JST), Tokyo, Japan

    Toshiaki Isobe

Authors
  1. Hiroyuki Kaji
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  2. Toshiaki Isobe
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Corresponding author

Correspondence to Hiroyuki Kaji .

Editor information

Editors and Affiliations

  1. Medical Center, Department of Biochemistry, University of Texas Southwestern, Harry Hines Blvd. 5323, Dallas, 75390-9038, Texas, USA

    Jennifer J. Kohler

  2. Medical Center, Dallas, University of Texas Southwestern, Harry Hines Blvd. 5323, Dallas, 75390-9185, Texas, USA

    Steven M. Patrie

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Kaji, H., Isobe, T. (2013). Stable Isotope Labeling of N-Glycosylated Peptides by Enzymatic Deglycosylation for Mass Spectrometry-Based Glycoproteomics. In: Kohler, J., Patrie, S. (eds) Mass Spectrometry of Glycoproteins. Methods in Molecular Biology, vol 951. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-146-2_14

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  • DOI: https://doi.org/10.1007/978-1-62703-146-2_14

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-145-5

  • Online ISBN: 978-1-62703-146-2

  • eBook Packages: Springer Protocols

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