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Glycoproteins Enrichment and LC-MS/MS Glycoproteomics in Central Nervous System Applications

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Neuroproteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1598))

Abstract

Proteins and glycoproteins play important biological roles in central nervous systems (CNS). Qualitative and quantitative evaluation of proteins and glycoproteins expression in CNS is critical to reveal the inherent biomolecular mechanism of CNS diseases. This chapter describes proteomic and glycoproteomic approaches based on liquid chromatography/tandem mass spectrometry (LC-MS or LC-MS/MS) for the qualitative and quantitative assessment of proteins and glycoproteins expressed in CNS. Proteins and glycoproteins, extracted by a mass spectrometry friendly surfactant from CNS samples, were subjected to enzymatic (tryptic) digestion and three down-stream analyses: (1) a nano LC system coupled with a high-resolution MS instrument to achieve qualitative proteomic profile, (2) a nano LC system combined with a triple quadrupole MS to quantify identified proteins, and (3) glycoprotein enrichment prior to LC-MS/MS analysis. Enrichment techniques can be applied to improve coverage of low abundant glycopeptides/glycoproteins. An example described in this chapter is hydrophilic interaction liquid chromatographic (HILIC) enrichment to capture glycopeptides, allowing efficient removal of peptides. The combination of three LC-MS/MS-based approaches is capable of the investigation of large-scale proteins and glycoproteins from CNS with an in-depth coverage, thus offering a full view of proteins and glycoproteins changes in CNS.

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Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Texas Tech University, Memorial Circle & Boston Ave., Box 41061, Lubbock, TX, 79409-1061, USA

    Rui Zhu, Ehwang Song & Yehia Mechref

  2. Department of Biotechnology, Institute of Graduate Studies and Research, University of Alexandria, Alexandria, 21526, Egypt

    Ahmed Hussein

  3. Department of Biochemistry and Molecular Genetics, Faculty of Medicine, American University of Beirut, Beirut, Lebanon

    Firas H. Kobeissy

  4. Department of Psychiatry, Center for Neuroproteomics and Biomarkers Research, University of Florida, Gainesville, FL, USA

    Firas H. Kobeissy

Authors
  1. Rui Zhu
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  2. Ehwang Song
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  3. Ahmed Hussein
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  4. Firas H. Kobeissy
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  5. Yehia Mechref
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Corresponding author

Correspondence to Yehia Mechref .

Editor information

Editors and Affiliations

  1. Department of Psychiatry, University of Florida McKnight Brain Institute, Gainesville, Florida, USA

    Firas H. Kobeissy

  2. Department of Cell Biology, Microbiology, & Molecular Biology, University of South Florida, Tampa, Florida, USA

    Stanley M. Stevens, Jr.

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Zhu, R., Song, E., Hussein, A., Kobeissy, F.H., Mechref, Y. (2017). Glycoproteins Enrichment and LC-MS/MS Glycoproteomics in Central Nervous System Applications. In: Kobeissy, F., Stevens, Jr., S. (eds) Neuroproteomics. Methods in Molecular Biology, vol 1598. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6952-4_9

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  • DOI: https://doi.org/10.1007/978-1-4939-6952-4_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6950-0

  • Online ISBN: 978-1-4939-6952-4

  • eBook Packages: Springer Protocols

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