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Structural Separations by Ion Mobility-MS for Glycomics and Glycoproteomics

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

Abstract

This chapter describes the utility of ion mobility-mass spectrometry (IM-MS) for the detection and characterization of glycoproteins and associated glycoconjugates. IM-MS provides separations in two dimensions; one on the basis of molecular surface area or structure, and the other on molecular mass which creates the ability to differentiate biomolecular classes and isobaric species. When applied to the characterization of glycoproteins, IM-MS separates peptides from the associated glycans in the same digest without purification, and can also be used to separate different isomeric glycans which is a significant challenge in current glycomic studies. The chapter details the methodologies to use IM-MS for the study of glycans and glycoproteins for an audience ranging from new and potential practitioners to those already utilizing the technique.

Key words

  • Ion mobility
  • Ion mobility-mass spectrometry
  • IM-MS
  • Structural separations
  • MALDI
  • IM-MS/MS
  • Glycomics
  • Glycoproteomics

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Acknowledgements

Financial support for this work was provided by the National Institutes of Health (1R01GM092218-01 and RC2DA028981), the US Defense Threat Reduction Agency (HDTRA-09-1-0013), Vanderbilt University College of Arts and Sciences, Vanderbilt Institute of Chemical Biology, and Vanderbilt Institute for Integrative Biosystems Research and Engineering. We thank Nichole M. Lareau for critical comments and assistance with this manuscript. Richard M. Caprioli (Vanderbilt University, Department of Biochemistry) and the Vanderbilt University Mass Spectrometry Research Center for use of the Synapt HDMS. The carbohydrate compounds, LacNAc, Lec, Lac, B tetra type 1, 2′F-B type 2, LNT, LeALex, Di-Lec, Di-LeA, LNnT, Galα3-type1, B2-tri, Pk, P1 tri, Tri-LacNAc, H-type2-LN-LN, P1 penta, P1 antigen, GNLNLN, and 3′SLN-Lec, were provided by the Carbohydrate Synthesis/Protein Expression Core of The Consortium for Functional Glycomics funded by the National Institute of General Medical Sciences grant GM62116.

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Fenn, L.S., McLean, J.A. (2013). Structural Separations by Ion Mobility-MS for Glycomics and 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_12

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