Sequence analysis of highly sulfated, heparin-derived oligosaccharides using fast atom bombardment mass spectrometry

Mallis, Larry M.; Wang, Hui M.; Loganathan, Duraikkannu; Linhardt, Robert J.

doi:10.1021/ac00188a030

Cited by 80 publications

(40 citation statements)

References 50 publications

(26 reference statements)

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“…Mass spectrometric investigation of heparin-derived oligosaccharides poses a serious challenge because they have to be extensively purified and desalted for negative-ion fastatom-bombardment mass spectrometry. This methodology requires large amounts of material (10 nmol/pJ) and still results in partially sodiated anions of monosulfated disaccharides and polysulfated di-to octasaccharides, which contain up to 15 Na+ ions (20)(21)(22). Probably because of these difficulties, little mass spectrometric work concerning this biologically important class of compounds has been reported to date.…”

Section: Resultsmentioning

confidence: 99%

Mass spectrometric molecular-weight determination of highly acidic compounds of biological significance via their complexes with basic polypeptides.

Juhász

Biemann

1994

Proc. Natl. Acad. Sci. U.S.A.

141

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Highly acidic compounds that are difficult to Ionize by matrix-assisted laser desorption onization give excellent spectra when mixed with a basic peptide or protein to form a noncovalent complex. This phenomenon makes it possible to determine the molecular weights of polysulfated, -sulfonated, and -phosphorylated blomolecules such as cysteic acid-containing peptides, oligonucleotides, heparin-derived oligosaccharides, and suramin (a drug containing two trisulfonated naphthalene moieties). Peptides and small proteins rich in arginine were used as the basic components. The extent of complex formation correlates with the number of phosphate and sulfate groups in the acidic component and with the number of arginines in the basic component. Neither the acidic amino acid residue aspartic and glutamic acid nor the basic lysine and histidine contribute to complex formation. For oligonudeotides, histone H4 was found to be the best complexing agent investigated. The analytical utility of the complex formation is demonstrated by the molecular-mass determination of acidic compounds from 500 to 6000 Da at the picomole or sub-picomole level with an accuracy of ± 0.1% or better and by the absence of alkali cation adducts.The development of matrix-assisted laser desorption ionization (MALDI) mass spectrometry permits the determination of the molecular mass of proteins up to the 105-Da range with an accuracy of 0.1-0.01%, requiring only picomoles or subpicomoles of material (1-4). The method is equally applicable to smaller biologically important molecules, such as peptides (5), carbohydrates (6), oligonucleotides (7,8), glycolipids (9), and polar and nonpolar synthetic polymers (10, 11). It has become an important technique in biochemistry and biology, not only because the molecular weight of the native material at that level of accuracy is in itself very useful information, but also because the changes thereof upon chemical or enzymatic treatment provide further insight into the structure or biological significance of parts of the native molecule (12). These manipulations are often necessary to obtain structural information because little excess energy is transferred to the analyte during the MALDI process, and "prompt" fragmentation is therefore rarely observed. This feature is an advantage in the analyses of mixtures, as long as the components can be resolved.Although most of the compounds in the above-mentioned categories are amenable to MALDI, it is difficult to ionize highly acidic compounds, even in the negative mode of the mass spectrometer, where they are detected as anions. In that mode, most efforts have been devoted to oligonucleotides (7,8). It is even more difficult to ionize polysulfate esters or polysulfonic acids. This is due, in part, to the fact that these substances tenaciously attach cations (such as Na+, K+, etc.) that give rise to broad unresolved peaks, the centroid of which corresponds to the average mass of all these partial salts.When attempting to use the oxidized A chain of bovine insulin (A...

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Section: Resultsmentioning

confidence: 99%

Mass spectrometric molecular-weight determination of highly acidic compounds of biological significance via their complexes with basic polypeptides.

Juhász

Biemann

1994

Proc. Natl. Acad. Sci. U.S.A.

141

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“…Methodologies using fast atom bombardment (FAB) were first employed; however a direct coupling to HPLC separation was not achieved, as in the case of peptides, due to limitations in mobile phase composition (Dell et al, 1988;Mallis et al, 1989). The introduction of soft ionization methods (ESI), permitting an easy interfacing to HPLC has first provided an online separation and structure elucidation of complex oligosaccharide mixtures (Henriksen et al, 2006;Barosso et al, 2005;Thanawiroon et al, 2004).…”

Section: Overview On Mass-spectrometric Analysis Of Gagsmentioning

confidence: 99%

HPLC-MS/MS of Highly Polar Compounds

Silvestro¹,

Tarcomnicu²,

Savu³

2012

Tandem Mass Spectrometry - Applications and Principles

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“…Characterization of heparin, HS, and other GAGs has been performed by fast atom bombardment (FAB) [7][8][9][10] and electrospray ionization (ESI) [11][12][13][14]. Different approaches using LC-ESI and tandem mass spectrometry (MS n ) were designed to differentiate isomeric heparin disaccharides [15], and determine sulfation positions and uronic acid epimerization in chondroitin sulfate oligosaccharides [16], as well as to study the effect of the positions of sulfate groups on heparin binding [2,17].…”

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confidence: 99%

UV-MALDI-TOF mass spectrometry analysis of heparin oligosaccharides obtained by nitrous acid controlled degradation and high performance anion exchange chromatography

Bultel

Landoni

Grand

et al. 2010

J. Am. Soc. Mass Spectrom.

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Nitrous acid degradation of heparin followed by high-performance anion-exchange chromatography (HPAEC) separation and ultraviolet matrix assisted laser desorption/ionization time-of-flight (UV-MALDI-TOF) analysis led to the structural determination of six sulfated oligosaccharides. Three different matrices (␣-cyano-4-hydroxycinnamic acid (CHCA), norharmane, and dihydroxybenzoic acid (DHB)) have been used, and the complementary results obtained allowed in most cases to assign the position of sulfate groups. Based on the different cleavages produced on the purified oligosaccharides in source during the MS analysis by the use of the different matrices, this approach provides a new tool for structural analysis. (J Am Soc Mass Spectrom 2010, 21, 178 -190) © 2010 American Society for Mass Spectrometry G lycosaminoglycanes (GAGs), and more particularly heparan sulfates (HS), have been implicated in the regulation of a broad range of biological processes because of their specific interactions with a large number of proteins [1]. These interactions seem to depend on the number and position of sulfate groups in individual sugar moieties and on the way that sulfated glycosidic units are regrouped to form domains [2,3].The basic structure of heparin or heparan-like glycosaminoglycans is a disaccharide comprising an uronic acid residue linked 1¡4 to a glucosamine. It is repeated many times to form heterogeneous mixtures of chains with different lengths. Various modifications within the basic disaccharide unit, including two possible epimers of the uronic acid (l-iduronic and d-glucuronic), sulfation at the N, 3-O, 6-O position of the glucosamine and 2-O position of the uronic acid, generate a considerable amount of diversity.Among the existing strategies used in current glycobiology to study HS, genetic approaches have shown that targeting a particular sulfotransferase gene may perturb many sulfation patterns throughout the polysaccharide chain and therefore cannot be used to study the impact of a single structural feature [4]. On the other hand, a number of different chemical and biological approaches have led to the generation of size-defined heparin oligosaccharide libraries [5,6]. However their complete purification and the sequencing analytical and chemical methods used for their characterization are still extremely laborious, complex, and particularly limited by the small amounts of purified products available, compromising the efforts to relate a biological function to a specific sulfation sequence. The limited amounts of pure oligosaccharide fragments obtained also preclude structural characterization by the use of spectroscopic methods such as NMR.The availability of "soft ionization" techniques pointed to MS as an attractive and prominent analytical technique for structural elucidation. Characterization of heparin, HS, and other GAGs has been performed by fast atom bombardment (FAB) [7][8][9][10] and electrospray ionization (ESI) [11][12][13][14]. Different approaches using LC-ESI and tandem mass spectrometry (MS ...

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Sequence analysis of highly sulfated, heparin-derived oligosaccharides using fast atom bombardment mass spectrometry

Cited by 80 publications

References 50 publications

Mass spectrometric molecular-weight determination of highly acidic compounds of biological significance via their complexes with basic polypeptides.

Mass spectrometric molecular-weight determination of highly acidic compounds of biological significance via their complexes with basic polypeptides.

HPLC-MS/MS of Highly Polar Compounds

UV-MALDI-TOF mass spectrometry analysis of heparin oligosaccharides obtained by nitrous acid controlled degradation and high performance anion exchange chromatography

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