Effect of monosaccharide composition, glycosidic linkage position and anomericity on the electrophoretic mobility of labeled oligosaccharides

Masuoka, James; Hazen, Kevin C.

doi:10.1002/elps.200500411

Cited by 3 publications

(2 citation statements)

References 29 publications

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“…5A, monosaccharide Gal cannot be separated from Gal-containing disaccharide lactose and moves faster than its other monosaccharide component Glc. Interestingly, disaccharide lactose (Gal(b 1-4)Glc) migrates significantly faster than its structural isomer melibiose (Gal(a 1-6)Glc), confirming the finding by other studies that in addition to charge, structural elements such as linkage position, and linkage anomericity contribute to glycan mobility [29]. The larger hydrodynamic radius and flexibility associated with the a 1-6 linkage could possibly explain the reduced electrophoretic mobility of melibiose, compared with lactose.…”

Section: Separation Of Mono-and Disaccharidessupporting

confidence: 85%

Boronate affinity saccharide electrophoresis: A novel carbohydrate analysis tool

et al. 2008

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The incorporation of specialised carbohydrate affinity ligand methacrylamido phenylboronic acid in polyacrylamide gels for fluorophore-assisted carbohydrate electrophoresis greatly improved the effective separation of saccharides that show similar mobilities in standard electrophoresis. Polyacrylamide gel electrophoresis using methacrylamido phenylboronic acid in low loading (typically 0.5-1% dry weight) was unequivocally shown to alter retention of labelled saccharides depending on their boronate affinity. While conventional fluorophore-assisted carbohydrate electrophoresis of 2-aminoacridone labelled glucose oligomers showed an inverted parabolic migration, an undesired trait of small oligosaccharides labelled with this neutral fluorophore, boron affinity saccharide electrophoresis separation of these carbohydrates completely restored their predicted running order, based on their charge/mass ratio, and resulted in improved separation of the analyte saccharides. These results exemplify boron affinity saccharide electrophoresis as an important new technique for analysing carbohydrates and sugar-containing molecules.

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Section: Separation Of Mono-and Disaccharidessupporting

confidence: 85%

Boronate affinity saccharide electrophoresis: A novel carbohydrate analysis tool

et al. 2008

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“…Recently, FACE has been proved to be useful not only for GAGs analysis but also for oligosaccharides from various sources, such as maltose and cellobiose [56]. For a better separation and more sensitive detection, these neutral saccharides were derivatized with the charged fluorophore ANTS.…”

Section: Fluorophores Name and Abbreviationmentioning

confidence: 99%

Analysis of Glycosaminoglycans by Electrophoretic Approach

Karousou¹,

Viola²,

Vigetti³

et al. 2008

CPA

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Glycosaminoglycans (GAGs) are non branched polysaccharides which are raising a great interest among the scientists for their biological roles. In fact GAGs play a pivotal role in several biological events, since they participate in and regulate cell adhesion, migration and proliferation. The quantification and analysis of the fine structure of GAGs are increasingly important not only for understanding many biological processes, but also for elucidate many critical aspects in human pathology development. Chondroitin sulfate (CS), heparan sulfate (HS) and keratan sulfate (KS) are commonly described as sulfated GAGs and these molecules are linked to a core protein forming proteoglycans; the sulfation pattern shows a high level of complexity and it is associated with specific function in the tissues. The only GAG without protein core is hyaluronan (HA), which is produced in almost all tissues, often with a molecular weight of 10 6 Daltons. Several human tissues contain high amount of GAGs and the change of the quantity and the structure of these macromolecules are described in tissue development and it is commonly associated with diseases. Electrophoretic methods based on the gel separation of 2-aminoacridone labelled HA and CS sulfate -disaccharides, derived from GAG digestion with specific eliminases, have been recently proposed. These new techniques represent a suitable method for GAG fast and sensitive analysis. In this review we will describe the recently achieved methods on the GAG analysis based on the electrophoretic approach in comparison with the more standard chromatographic techniques (HPLC).

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Influence of outer region mannosylphosphorylation on N-glycan formation by Candida albicans: Normal acid-stable N-glycan formation requires acid-labile mannosylphosphate addition

2007

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The pathogenic yeast Candida albicans produces large N-glycans with outer regions containing only mannose residues. The outer region comprises a primary branch with multiple secondary and tertiary branches. Tertiary branches are linked to secondary branches by phosphodiester bridges. In the current model of outer chain elongation in the genetically related yeast Saccharomyces cerevisiae, synthesis of the branches occurs sequentially, primary to tertiary. Thus, disruption of mannosylphosphorylation, the initial step in tertiary branch formation, should not affect primary or secondary branch production. Compared to its wild-type parent, a C. albicans mutant defective in tertiary branch mannosylphosphorylation (mnn4Delta/mnn4Delta) made outer regions with reduced susceptibility to low acid acetolysis treatment, suggesting that the secondary or primary region had been modified. Higher acid acetolysis conditions were required to release the secondary branches from the primary branches. The released secondary branches constitute the subset of the wild-type secondary branches that lack a phosphate group. In contrast, the acid-stable region of both wild-type and mnn4Delta S. cerevisiae strains required high acid acetolysis conditions to release the secondary branches, despite having smaller and less complex secondary and tertiary branches. These results suggest that the complex and longer secondary and tertiary branches of C. albicans affect the conformation of the acid-stable region to render it more susceptible to acetolysis which implies secondary and tertiary branch formation in C. albicans are interdependent events and occur concurrently, rather than sequentially.

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Effect of monosaccharide composition, glycosidic linkage position and anomericity on the electrophoretic mobility of labeled oligosaccharides

Cited by 3 publications

References 29 publications

Boronate affinity saccharide electrophoresis: A novel carbohydrate analysis tool

Boronate affinity saccharide electrophoresis: A novel carbohydrate analysis tool

Analysis of Glycosaminoglycans by Electrophoretic Approach

Influence of outer region mannosylphosphorylation on N-glycan formation by Candida albicans: Normal acid-stable N-glycan formation requires acid-labile mannosylphosphate addition

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