Primary structure of seven sulfated oligosaccharide-alditols released by reductive β-elimination from oviducal mucins of Rana temporaria characterization of the sequence HSO3(3)GlcA(β1–3)Gal

Florea, Doina; Maës, Emmanuel; Strecker, Gérard

doi:10.1016/s0008-6215(97)00129-8

Cited by 19 publications

(20 citation statements)

References 21 publications

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“…The NMR parameters (Table 2) of this unit are typical of the (α1–2) linkage to Gal [25]. On the basis of our previous work, we suspected that the hexuronic acid was a GlcA unit, sometimes found in other amphibian carbohydrate molecules[9,13,14]. After carboxymethylation, reduction with NaBD 4 , permethylation, methanolysis and acetylation, the GC/MS analysis showed the presence of 6,6′‐D 2 ‐(2,3,4,6)‐Me 4 ‐Glc, showing the existence of GlcA in nonreducing terminal position.…”

Section: Resultsmentioning

confidence: 99%

“…and H‐4 (Δδ = +0.370 p.p.m.) are attributable to a C‐ 3 substitution with a sulfate group [14]. Furthermore, the ESI‐MS/MS analysis (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…All the collected fractions were desalted by gel permeation on a Bio‐Gel P2 column (55 × 2 cm), and lyophilized [20]. The different steps of the purification are explained in detail in many previous papers [12–17].…”

Section: Methodsmentioning

confidence: 99%

“…Previous studies have shown a species‐specific structural diversity in carbohydrate chains of amphibian egg jelly coats [8–19]. Consequently, these carbohydrate chains represent new phenotypic markers of amphibian species and from a biological point of view, can be involved in species‐specific interaction gametes or in the specific host–parasite interaction [20].…”

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confidence: 99%

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Acquisition of species‐specific O‐linked carbohydrate chains from oviducal mucins inRana arvalis

Coppin

Maës

Flahaut

et al. 1999

European Journal of Biochemistry

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The extracellular matrix surrounding amphibian eggs is composed of mucin-type glycoproteins, highly O-glycosylated and plays an important role in the fertilization process. Oligosaccharide-alditols were released from the oviducal mucins of the anuran Rana arvalis by alkali-borohydride treatment in reduced conditions. Neutral and acidic oligosaccharides were fractionated by ion-exchange chromatographies and purified by HPLC. Each compound was identified by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) spectrometry, NMR spectroscopy, electrospray ionization-tandem mass spectroscopy (ESI-MS/MS) and permethylation analyses. This paper reports on the structures of 19 oligosaccharide-alditols, 12 of which have novel structures. These structures range in size from disaccharide to octasaccharide. Some of them are acidic, containing either a glucuronic acid or, more frequently, a sulfate group, located either at the 6 position of GlcNAc or the 3 or 4 positions of Gal. This latter sulfation is novel and has only been characterized in the species R. arvalis. This structural analysis led to the establishment of several novel carbohydrate structures, demonstrating the structural diversity and species-specificity of amphibian glycoconjugates.Keywords: sulfated-oligosaccharides; NMR; amphibian; oviducal mucin.The amphibian eggs are surrounded by oviducal jellies, which can be structurally and chemically distinct from between species [1]. The jelly coats are made up of several concentric layers composed of mucin-type glycoproteins, highly O-glycosylated. These glycoproteins have been shown to play important roles in fertilization [2,3]. Many functions have been assigned to the egg jelly components such as sperm binding, sperm capacitation, induction of the acrosome reaction, prevention of species cross-fertilization, the block of polyspermy (anurans) and provision of a protective ionic environment for the developing embryo [4±7].Previous studies have shown a species-specific structural diversity in carbohydrate chains of amphibian egg jelly coats [8±19]. Consequently, these carbohydrate chains represent new phenotypic markers of amphibian species and from a biological point of view, can be involved in species-specific interaction gametes or in the specific host±parasite interaction [20]. In the present study, we have isolated neutral and acidic carbohydrate chains from the oviducal mucins of Rana arvalis, and determined the structure of 19 of them. M A T E R I A L S A N D M E T H O D S Sampling of jelly coat mucusEggs from R. arvalis, collected in France and Russia, were obtained from natural spawns. The jelly coat material was lyophilized. Isolation of oligosaccharide-alditolsO-linked oligosaccharides were released from the crude material by alkaline degradation in reduced conditions (1 m NaBH 4 ) to obtain stable carbohydrate chains. Total material was submitted to a cationic exchange chromatography on Dowex 50 Â 2 (200±400 mesh, H + form) to remove residual peptides. Neutral oligosaccharide-alditols ...

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Section: Resultsmentioning

confidence: 99%

“…and H‐4 (Δδ = +0.370 p.p.m.) are attributable to a C‐ 3 substitution with a sulfate group [14]. Furthermore, the ESI‐MS/MS analysis (Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Acquisition of species‐specific O‐linked carbohydrate chains from oviducal mucins inRana arvalis

Coppin

Maës

Flahaut

et al. 1999

European Journal of Biochemistry

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“…We recently identified a 3-O-sulfotransferase from ovarian, breast, and other tissues capable of incorporating sulfate esters at C-3 of Gal in the Galβ1-3GalNAc sequence (Kuhns et al, 1995;Chandrasekaran et al, 1996. Recently, reports of a native oligosaccharide carrying SE-3Galβ1-3Gal-NAc have appeared (Chance and Mawhinney, 1996;Florea et al, 1997). We synthesized a branched derivative Core 2 structure 8, containing a 3-O-sulfate ester group instead of sialic acid moiety and found it to be inhibitory for L-and P-selectin (Koenig et al, 1997).…”

Section: Inhibition Studiesmentioning

confidence: 99%

Inhibition of L- and P-selectin by a rationally synthesized novel core 2-like branched structure containing GalNAc-Lewisx and Neu5Ac 2-3Gal 1-3GalNAc sequences

Jain¹,

Piskorz

Huang

et al. 1998

Glycobiology

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The selectins interact in important normal and pathological situations with certain sialylated, fucosylated glycoconjugate ligands containing sialyl Lewis x (Neu5Acα2-3Galβ1-4(Fuc-α1-3)GlcNAc). Much effort has gone into the synthesis of sialylated and sulfated Lewis x analogs as competitive ligands for the selectins. Since the natural selectin ligands GlyCAM-1 and PSGL-1 carry sialyl Lewis x as part of a branched Core 2 O-linked structure, we recently synthesized Galβ1-4(Fuc-α1-3)GlcNAcβ1-6(SE-3Galβ1-3)GalNAc1αOMe and found it to be a moderately superior ligand for L and P-selectin (Koenig et al., Glycobiology 7, 79-93, 1997). Other studies have shown that sulfate esters can replace sialic acid in some selectin ligands (Yeun et al., Biochemistry, 31, 9126-9131, 1992; Imai et al., Nature, 361, 555, 1993). Based upon these observations, we hypothesized that Neu5Acα2-3Galβ1-3Gal-NAc might have the capability of interacting with L-and P-selectin. To examine this hypothesis, we synthesized Galβ1-4(Fucα1-3)GlcNAcβ1-6(Neu5Acα2-3Galβ1-3)-GalNAcα1-OB, which was found to be 2-to 3-fold better than sialyl Le x for P and L selectin, respectively. We also report the synthesis of an unusual structure GalNAcβ1-4(Fucα1-3)GlcNAcβ1-OMe (GalNAc-Lewis x -O-methyl glycoside), which also proved to be a better inhibitor of L-and P-selectin than sialyl Lewis x -OMe. Combining this with our knowledge of Core 2 branched structures, we have synthesized a molecule that is 5-to 6-fold better at inhibiting L-and P-selectin than sialyl Lewis x -OMe, By contrast to unbranched structures, substitution of a sulfate ester group for a sialic acid residue in such a molecule resulted in a considerable loss of inhibition ability. Thus, the combination of a sialic acid residue on the primary (β1-3) arm, and a modified Le x unit on the branched (β1-6) arm on an O-linked Core 2 structure generated a monovalent synthetic oliogosaccharide inhibitor superior to SLe x for both L-and P-selectin.

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Fragmentations of isomeric sulfated monosaccharides using electrospray ion trap mass spectrometry

Minamisawa

Hirabayashi

2005

Rapid Comm Mass Spectrometry

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Electrospray ionization combined with ion trap mass spectrometry (ESI-ITMS) is a powerful tool for structural analysis of complex carbohydrates. Although its application to sulfated glycans has been limited so far, it should provide critical information, such as sulfate positions, on their structures. In this work, MS(n) spectra of nine monosulfated monosaccharides, consisting of five hexoses and four N-acetylhexosamines, were measured in negative ion mode to find basic fragmentation rules for sulfated sugars. Two pairs of positional isomers with respect to sulfation, i.e., Gal4S and Gal6S, and GalNAc4S and GalNAc6S, showed characteristic fragmentation patterns in MS(3), and could be discriminated from one another by the appearance of particular diagnostic fragment ions that characterize individual isomers. It was also demonstrated that, even if a mixture of these positional isomers was analyzed, the proportion of each species could be estimated through analysis of the abundance ratios of the diagnostic ions. However, 3-O-sulfated saccharides (Glc3S and GlcNAc3S) gave a single abundant diagnostic ion in MS(2) corresponding to the hydrogensulfate ion, [OSO(3)H](-), and this characteristic clearly differentiated them from their positional isomers. In contrast, 6-O-sulfated diastereomers consisting of two groups, Glc6S, Man6S, Gal6S, and GlcNAc6S, GalNAc6S, could not be discriminated by the types of fragment ions; however, the abundance ratios of particular fragment ions differed significantly between Glc(NAc)6S and Gal(NAc)6S. Since ESI-ITMS yielded large quantities of useful information on structures of monosulfated hexoses and N-acetylhexosamines in an extremely simple and reproducible manner, establishment of a comprehensive strategy based on ESI-ITMS(n) appears to be a promising technique for structural elucidation of sulfated complex carbohydrates.

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Primary structure of seven sulfated oligosaccharide-alditols released by reductive β-elimination from oviducal mucins of Rana temporaria characterization of the sequence HSO3(3)GlcA(β1–3)Gal

Cited by 19 publications

References 21 publications

Acquisition of species‐specific O‐linked carbohydrate chains from oviducal mucins inRana arvalis

Acquisition of species‐specific O‐linked carbohydrate chains from oviducal mucins inRana arvalis

Inhibition of L- and P-selectin by a rationally synthesized novel core 2-like branched structure containing GalNAc-Lewisx and Neu5Ac 2-3Gal 1-3GalNAc sequences

Fragmentations of isomeric sulfated monosaccharides using electrospray ion trap mass spectrometry

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