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Glycobiology Advance Access originally published online on July 10, 2007
Glycobiology 2007 17(9):972-982; doi:10.1093/glycob/cwm072
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© The Author 2007. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

Towards GAG glycomics: Analysis of highly sulfated heparins by MALDI-TOF mass spectrometry

Bérangère Tissot2, Nijole Gasiunas3, Andrew K Powell4, Yassir Ahmed4, Zheng-liang Zhi4, Stuart M Haslam2, Howard R Morris2,5,, Jeremy E Turnbull4, John T Gallagher3 and Anne Dell1,2

2 Division of Molecular Biosciences, Imperial College, London SW7 2AZ, UK
3 Cancer Research UK, Department of Medical Oncology, University of Manchester, Christie Hospital, Manchester, M20 4BX, UK
4 Molecular Glycobiology Laboratory, School of Biological Sciences, University of Liverpool, L69 7ZB, UK
5 M-SCAN Ltd, Wokingham, Berks, RG41 2TZ, UK

1 To whom correspondence should be addressed: Tel: + 44-207-5945219; Fax: + 44-207-2250458; e-mail: a.dell{at}imperial.ac.uk

Received on April 22, 2007; revised on June 26, 2007; accepted on June 27, 2007

Glycomics is a developing field that provides structural information on complex populations of glycans isolated from tissues, cells and organs. Strategies employing matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) are central to glycomic analysis. Current MALDI-based glycomic strategies are capable of efficiently analyzing glycoprotein and glycosphingolipid glycomes but little attention has been paid to devising glycomic methodologies suited to the analysis of glycosaminoglycan (GAG) polysaccharides which pose special problems for MALDI analysis because of their high level of sulfation and large size. In this paper, we describe MALDI strategies that have been optimized for the analysis of highly sulfated GAG-derived oligosaccharides. A crystalline matrix norharmane, as well as an ionic liquid 1-methylimidazolium {alpha}-cyano-4-hydroxycinnamate (ImCHCA), have been used for the analysis of heparin di-, tetra-, hexa- and decasaccharides carrying from 2 to 13 sulfate groups. Information about the maximum number of sulfate groups is obtained using the ionic liquid whereas MALDI-TOF/TOF MS/MS experiments using norharmane allowed the determination of the nature of the glycosidic backbone, and more precise information about the presence and the position in the sequence of N-acetylated residues.

Key words: glycomics / glycosaminoglycans / ionic liquid / MALDI mass spectrometry


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