Analysis of Glycosaminoglycans Using Mass Spectrometry

Abstract

The glycosaminoglycans (GAGs) are linear polysaccharides expressed on animal cell surfaces and in extracellular matrices. Their biosynthesis is under complex control and confers a domain structure that is essential to their ability to bind to protein partners. Key to understanding the functions of GAGs are methods to determine accurately and rapidly patterns of sulfation, acetylation and uronic acid epimerization that correlate with protein binding or other biological activities. Mass spectrometry (MS) is particularly suitable for the analysis of GAGs for biomedical purposes. Using modern ionization techniques it is possible to accurately determine molecular weights of GAG oligosaccharides and their distributions within a mixture. Methods for direct interfacing with liquid chromatography have been developed to permit online mass spectrometric analysis of GAGs. New tandem mass spectrometric methods for fine structure determination of GAGs are emerging. This review summarizes MS-based approaches for analysis of GAGs, including tissue extraction and chromatographic methods compatible with LC/MS and tandem MS.

Isomer separation and effect of the degree of polymerization on the gas‐phase structure of chondroitin sulfate oligosaccharides analyzed by ion mobility and tandem mass spectrometry

“…Mass spectrometry (MS) using the soft ionization technique electrospray (ESI) has been widely recognized as a powerful and highly sensitive method for the structural analysis of sulfated GAGs . The fine structural analysis of GAGs by MS is most commonly carried out on GAG fragments, mainly at the disaccharide (dp2) level, after the enzymatic or chemical depolymerization of the GAG chain.…”

Effects of calcium complexation on heparin‐like disaccharides. A combined theoretical, tandem mass spectrometry and ultraviolet experiment

“…[5,14] Structural characterization of sulfated Hp benefited from the development of mass spectrometry techniques applied to oligosaccharides. [19][20][21][22][23][24][25][26][27] Nevertheless, the fragmentation behavior of sulfated Hps has proven difficult to analyze. Sulfate group losses prevent full structural characterization by collisioninduced dissociation (CID).…”

Identification of an unusually sulfated tetrasaccharide chondroitin/dermatan motif in mouse brain by combining chip‐nanoelectrospray multistage MS2‐MS4 and high resolution MS

“…This cleavage is independent of sulfation pattern and content of GalNAc and GlcA [56]; therefore all GalNAc-GlcA sequences represent substrates prone to be cleaved by the enzyme, even under nonexhaustive digestion conditions. In view of this aspect, demonstrated previously [19,21,25,31,48,49], it appears obvious that the chains longer than disaccharide contain GalNAc-IdoA linkages for which Chondroitinase AC I Flavo has no specificity. Even if the stereochemistry of the hexuronic acid from the nonreducing end was lost, for a better insight into the structure of these complex molecules we kept the original annotation due to the known origin of GlcA.…”

“…Structural determination of CS/DS domains is a crucial step in the assessment of the biological role played by these molecules at the CNS level. For CS/DS compositional analysis and sequence correlation with its biological activity, MS was shown lately to be one the most proficient method [13][14][15][16][17][18][19][20][21]. MS applications in GAG analysis expanded after the optimization of the ESI [15,16,[22][23][24][25][26] and MALDI [27][28][29][30] methods as well as of multistage MS (MS n ) by CID [21,31] and electron-detachment dissociation (EDD) [32][33][34] for sequencing of single molecular species in complex mixtures.…”