Ion-spray mass spectrometry for identification of the nonreducing terminal sugar of glycosaminoglycan

Takagaki, Keiichi; Munakata, Hidekazu; Nakamura, Wataru; Matsuya, Hideki; Majima, Mitsuo; Endo, Masahiko

doi:10.1093/glycob/8.7.719

Cited by 14 publications

(4 citation statements)

References 26 publications

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“…GlcA␤1-3GalNAc(4S,6S)␤1-4GlcA␤1-3GalNAc(4S,6S)␤1-4GlcA␤1-3GalNAc(4S,6S)␤1-4GlcA was purified with a Bio-Gel P-4 column after ␤-elimination of GlcA␤1-3GalNAc(4S,6S)␤1-4GlcA␤1-3GalNAc-(4S,6S)␤1-4GlcA␤1-3GalNAc(4S,6S)␤1-4GlcA␤1-3GalNAc(4S,6S). The ␤-elimination reaction was carried out by incubation in a saturated solution of Ca(OH) 2 at 25°C for 4 h as described previously (19).…”

Section: Methodsmentioning

confidence: 99%

“…Ion-spray Mass Spectrometry-Mass spectra were obtained on an API-100 triple-quadruple mass spectrometer (PE SCIEX, Ontario, Canada) equipped with an atmospheric pressure ionization source, as described previously (18,19). The samples were dissolved in 0.5 mM ammonium acetate-acetonitrile (50:50) and injected at 2 l/min with a micro-HPLC syringe pump (Pump 22; Harvard Apparatus).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Domain Structure of Chondroitin Sulfate E Octasaccharides Binding to Type V Collagen

Takagaki

Munakata

Kakizaki

et al. 2002

Journal of Biological Chemistry

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We demonstrated previously that chondroitin sulfate E (ChS-E) binds to type V collagen (Munakata, H., Takagaki, K., Majima, M., and Endo, M. (1999) Glycobiology 9, 1023-1027). In this study, we investigated the structure and binding of ChS-E oligosaccharides. Eleven oligosaccharides were isolated from ChS-E by gel filtration chromatography and anion-exchange high performance liquid chromatography after hydrolysis with testicular hyaluronidase. Separately, seven oligosaccharides were custom synthesized using the transglycosylation reaction of testicular hyaluronidase. Structural analysis was performed by enzymatic digestions in conjunction with high performance liquid chromatography and mass spectrometry. This library of 18 oligosaccharides was used as a source of model molecules to clarify the structural requirements for binding to type V collagen. Binding was analyzed by a biosensor based on surface plasmon resonance. The results indicated that to bind to type V collagen the oligosaccharides must have the following carbohydrate structures: 1) octasaccharide or larger in size; 2) a continuous sequence of three GlcA␤1-3GalNAc(4S,6S) units; 3) a GlcA␤1-3GalNAc(4S,6S) unit, GlcA␤1-3GalNAc(4S) unit or GlcA␤1-3GalNAc(6S) unit at the reducing terminal; 4) a GlcA␤1-3GalNAc(4S,6S) unit at the nonreducing terminal. It is likely that these characteristic oligosaccharide sequences play key roles in cell adhesion and extracellular matrix assembly.Proteoglycan exists in tissues of many mammalian species and is widely distributed in the cell surface and extracellular matrix. It is known that its carbohydrate chain, chondroitin sulfate (ChS), 1 has various important biological activities in areas such as cell migration, recognition, and morphogenesis (1-5). Recently, greater attention has been directed toward the functions of ChS in the brain, optic nerves, and chondrocytes (6 -8). ChS has many types of structural domains that are known to participate in specific physiological functions. However, the relationship between the biological function and structure of ChS domains is not yet fully understood. Recently, we used a surface plasmon resonance (SPR) biosensor to investigate the interaction of glycosaminoglycans (GAGs) with collagens and glycoproteins from the extracellular matrix (9). It was found that chondroitin sulfate E (ChS-E) has specific affinity for type V collagen, and that a novel characteristic sequence included in ChS-E is probably involved in binding to type V collagen. ChS-E from squid cartilage consists mainly of the disaccharide units GlcA␤1-3GalNAc(4S,6S), GlcA␤1-3GalNAc(4S), GlcA␤1-3GalNAc(6S), and GlcA␤1-3GalNAc, where 4S and 6S represent 4-O-and 6-O-sulfate, respectively (10, 11). Thus, since ChS-E has a variety of structures, it is likely that some specific feature of its structure is necessary for binding to type V collagen. However, the structural features within ChS-E that bind to type V collagen remain unclear.In the present work, we have prepared various oligosaccharides by testicular hyaluronidase...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Domain Structure of Chondroitin Sulfate E Octasaccharides Binding to Type V Collagen

Takagaki

Munakata

Kakizaki

et al. 2002

Journal of Biological Chemistry

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“…MALDI time-of-flight (TOF) MS (19,26) and 23,33,34) have been the most favored approaches for the analysis of GAG-derived oligosaccharides. Both ESI-MS and MALDI-TOF-MS enjoy particular advantages in the analysis of large polar macromolecules.…”

Section: Liquid Chromatography/mass Spectrometry (Lc/ms)mentioning

confidence: 99%

Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides

Thanawiroon

Rice

Toida

et al. 2004

Journal of Biological Chemistry

146

152

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Liquid chromatography/mass spectrometry (LC/MS)is applied to the analysis of complex mixtures of oligosaccharides obtained through the controlled, heparinase-catalyzed depolymerization of heparin. Reversedphase ion-pairing chromatography, utilizing a volatile mobile phase, results in the high resolution separation of highly sulfated, heparin-derived oligosaccharides. Simultaneous detection by UV absorbance and electrospray ionization-mass spectrometry (ESI-MS) provides important structural information on the oligosaccharide components of this mixture. Highly sensitive and easily interpretable spectra were obtained through post-column addition of tributylamine in acetonitrile. High resolution mass spectrometry afforded elemental composition of many known and previously unknown heparin-derived oligosaccharides. UV in combination with MS detection led to the identification of oligosaccharides arising from the original non-reducing end (NRE) of the heparin chain. The structural identification of these oligosaccharides provided sequence from a reading frame that begins at the non-reducing terminus of the heparin chain. Interestingly, 16 NRE oligosaccharides are observed, having both an even and an odd number of saccharide residues, most of which are not predicted based on biosynthesis or known pathways of heparin catabolism. Quantification of these NRE oligosaccharides afforded a number-averaged molecular weight consistent with that expected for the pharmaceutical heparin used in this analysis. Molecular ions could be assigned for oligosaccharides as large as a tetradecasaccharide, having a mass of 4625 Da and a net charge of ؊32. Furthermore, MS detection was demonstrated for oligosaccharides with up to 30 saccharide units having a mass of >10,000 Da and a net charge of ؊60.The structural elucidation of complex carbohydrates remains one of the most difficult challenges for chemists, often requiring the application of multiple analytical approaches (1-5). Glycosaminoglycans (GAGs), 1 and heparin in particular, have proven to be extremely difficult to analyze because of high negative charge, polydispersity, and sequence heterogeneity (6, 7). Heparin and low molecular weight heparins, prepared through the controlled chemical or enzymatic fragmentation of heparin (8), are widely used as clinical anticoagulants. Despite their medical importance, these drugs are relatively uncharacterized in terms of their chemical structure. Moreover, heparin and the structurally related heparan sulfate exhibit many additional biological activities, making them of great interest in new drug discovery (9). Numerous challenges can arise from the structural investigation of biologically active heparin oligosaccharides, particularly those in recognition systems involving specific protein-carbohydrate interactions (10, 11). Such biologically important oligosaccharides often contain rare sequences (12, 13) and are present only in minute, often picomole quantities. New derivatization methods (14, 15), chromatography (15, 16), electrophoresis-ba...

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“…Soft ionization techniques, including ESI and matrix-assisted laser desorption/ionization (MALDI) are more dedicated to the analysis of neutral oligosaccharides and their attached peptide and lipid conjugates [29, 39]. MALDI time-of-flight (TOF) MS [40, 41] and ESI-MS [42] are useful approaches for the structural analysis of GAG-derived oligosaccharides. Both ESI-MS and MALDI-TOF-MS are convenient for structural analysis of large polar macromolecules.…”

Section: Introductionmentioning

confidence: 99%

Implementation of infrared and Raman modalities for glycosaminoglycan characterization in complex systems

et al. 2016

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Glycosaminoglycans (GAGs) are natural, linear and negatively charged heteropolysaccharides which are incident in every mammalian tissue. They consist of repeating disaccharide units, which are composed of either sulfated or non-sulfated monosaccharides. Depending on tissue types, GAGs exhibit structural heterogeneity such as the position and degree of sulfation or within their disaccharide units composition being heparin, heparan sulfate, chondroitine sulfate, dermatan sulfate, keratan sulfate, and hyaluronic acid. They are covalently linked to a core protein (proteoglycans) or as free chains (hyaluronan). GAGs affect cell properties and functions either by direct interaction with cell receptors or by sequestration of growth factors. These evidences of divert biological roles of GAGs make their characterization at cell and tissue levels of importance. Thus, non-invasive techniques are interesting to investigate, to qualitatively and quantitatively characterize GAGs in vitro in order to use them as diagnostic biomarkers and/or as therapeutic targets in several human diseases including cancer. Infrared and Raman microspectroscopies and imaging are sensitive enough to differentiate and classify GAG types and subtypes in spite of their close molecular structures. Spectroscopic markers characteristic of reference GAG molecules were identified. Beyond these investigations of the standard GAG spectral signature, infrared and Raman spectral signatures of GAG were searched in complex biological systems like cells. The aim of the present review is to describe the implementation of these complementary vibrational spectroscopy techniques, and to discuss their potentials, advantages and disadvantages for GAG analysis. In addition, this review presents new data as we show for the first time GAG infrared and Raman spectral signatures from conditioned media and live cells, respectively.

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Ion-spray mass spectrometry for identification of the nonreducing terminal sugar of glycosaminoglycan

Cited by 14 publications

References 26 publications

Domain Structure of Chondroitin Sulfate E Octasaccharides Binding to Type V Collagen

Domain Structure of Chondroitin Sulfate E Octasaccharides Binding to Type V Collagen

Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides

Implementation of infrared and Raman modalities for glycosaminoglycan characterization in complex systems

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