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Mass Spectrometry of Proteins and Peptides pp 181–200Cite as

Structure Analysis of N -Glycoproteins

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Part of the book series: Methods In Molecular Biology ((MIMB,volume 492))

Summary

General mass spectrometry-based strategies for analysis of N -glycosylated peptides are described. The well-established method utilizes Peptide-N -glycosidase F (PNGase F) for in-gel or in-solution release of N -linked glycans from the polypeptide chains (along with the conversion of the formerly N -glycosylated Asn to Asp), thus allowing separate analysis of glycan moieties and deglycosylated peptides. However, no assignment of individual glycans to a glycosylation site can be realized. Intact glycopeptides (i.e., proteolytic mixtures in which the glycan chains stay attached at their original glycosylation sites) can be analyzed either by a direct infusion or with HPLC separation prior to MALDI or ESI mass spectrometric analysis to provide both information on the glycan structure and glycosylation site in the same experiment. Several different strategies for efficient in-solution digestion of glycoproteins are described, such as proteolytic digestion in the electrospray capillary and simultaneous analysis of the resulting (glyco)peptides.

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Acknowledgments

We thank the “Deutsche Forschungsgemeinschaft” (SFB 492/Z2) for financial support.

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Authors and Affiliations

  1. Institute for Medical Physics and Biophysics, University of Münster, Münster, Germany

    Stefanie Henning, Jasna Peter-Katalinić & Gottfried Pohlentz

Authors
  1. Stefanie Henning
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  2. Jasna Peter-Katalinić
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  3. Gottfried Pohlentz
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Editor information

Editors and Affiliations

  1. Fundamental & Computational Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, USA

    Mary S. Lipton

  2. Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA

    Ljiljana Paša-Tolic

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Henning, S., Peter-Katalinić, J., Pohlentz, G. (2009). Structure Analysis of N -Glycoproteins. In: Lipton, M.S., Paša-Tolic, L. (eds) Mass Spectrometry of Proteins and Peptides. Methods In Molecular Biology, vol 492. Humana Press. https://doi.org/10.1007/978-1-59745-493-3_10

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  • DOI: https://doi.org/10.1007/978-1-59745-493-3_10

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-934115-48-0

  • Online ISBN: 978-1-59745-493-3

  • eBook Packages: Springer Protocols

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