Chromatographic separation of asparagine-linked oligosaccharides labeled with an ultravioletabsorbing compound,p-aminobenzoic acid ethyl ester

Matsuura, Fumito; Imaoka, Akiko

doi:10.1007/bf01048328

Cited by 43 publications

(30 citation statements)

References 27 publications

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“…S5). To confirm purity, b-1,3-Gal n and b-1,6-Gal n were coupled at their reducing terminals with p-aminobenzoic acid ethyl ester according to the method of Matsuura and Imaoka (1988) and analyzed on an HPLC system equipped with a TSKgel Amide-80 (4.6 3 250 mm; Tosoh) column ( Fig. 5; Supplemental Fig.…”

Section: Preparation Of Oligosaccharidesmentioning

confidence: 99%

β-Galactosyl Yariv Reagent Binds to the β-1,3-Galactan of Arabinogalactan Proteins

et al. 2013

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Yariv phenylglycosides [1,3,5-tri(p-glycosyloxyphenylazo)-2,4,6-trihydroxybenzene] are a group of chemical compounds that selectively bind to arabinogalactan proteins (AGPs), a type of plant proteoglycan. Yariv phenylglycosides are widely used as cytochemical reagents to perturb the molecular functions of AGPs as well as for the detection, quantification, purification, and staining of AGPs. However, the target structure in AGPs to which Yariv phenylglycosides bind has not been determined. Here, we identify the structural element of AGPs required for the interaction with Yariv phenylglycosides by stepwise trimming of the arabinogalactan moieties using combinations of specific glycoside hydrolases. Whereas the precipitation with Yariv phenylglycosides (Yariv reactivity) of radish (Raphanus sativus) root AGP was not reduced after enzyme treatment to remove a-L-arabinofuranosyl and b-glucuronosyl residues and b-1,6-galactan side chains, it was completely lost after degradation of the b-1,3-galactan main chains. In addition, Yariv reactivity of gum arabic, a commercial product of acacia (Acacia senegal) AGPs, increased rather than decreased during the repeated degradation of b-1,6-galactan side chains by Smith degradation. Among various oligosaccharides corresponding to partial structures of AGPs, b-1,3-galactooligosaccharides longer than b-1,3-galactoheptaose exhibited significant precipitation with Yariv in a radial diffusion assay on agar. A pull-down assay using oligosaccharides cross linked to hydrazine beads detected an interaction of b-1,3-galactooligosaccharides longer than b-1,3-galactopentaose with Yariv phenylglycoside. To the contrary, no interaction with Yariv was detected for b-1,6-galactooligosaccharides of any length. Therefore, we conclude that Yariv phenylglycosides should be considered specific binding reagents for b-1,3-galactan chains longer than five residues, and seven residues are sufficient for cross linking, leading to precipitation of the Yariv phenylglycosides.Arabinogalactan proteins (AGPs) are a type of plant proteoglycans consisting of a Hyp-rich core protein and large arabinogalactan (AG) moieties (Fincher et al., 1983;Nothnagel, 1997). Although there are many molecular species of AGP differentiated by their core proteins, the AG moieties commonly comprise b-1,3-galactan main chains and b-1,6-galactan side chains, to which L-Ara and other auxiliary sugars, such as GlcA, 4-O-methyl-GlcA, L-Fuc, L-Rha, and Xyl, are attached (Fincher et al., 1983;Nothnagel, 1997;Seifert and Roberts, 2007). A commercial product of AGPs prepared from the acacia (Acacia senegal) tree is known as gum arabic and utilized as a food stabilizer. In the Japanese herbal remedy Juzen-Taiho-To, AGs from Astragalus membranaceus are the active ingredient (Majewska-Sawka and Nothnagel, 2000;Kiyohara et al., 2002). In intact plants, AGPs are implicated in various physiological events and serve as extracellular constituents and signaling molecules. For instance, an AGP from stylar transmitting tissue attracts pollen tub...

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Section: Preparation Of Oligosaccharidesmentioning

confidence: 99%

β-Galactosyl Yariv Reagent Binds to the β-1,3-Galactan of Arabinogalactan Proteins

et al. 2013

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“…For the analysis of transglycosylation activity, 10 mM ␤-1,3-Gal 3 was used as the substrate. After incubation at 37°C, the reducing sugars in the reaction mixtures were coupled at their reducing terminals with p-aminobenzoic acid ethyl ester (ABEE) according to the method of Matsuura and Imaoka (36). The ABEE-derivatized sugars were analyzed on a highperformance liquid chromatography (HPLC) system equipped with a TSKgel Amide-80 (4.6 ϫ 250 mm; Tosoh, Tokyo, Japan) column as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

Endo-β-1,3-galactanase from Winter Mushroom Flammulina velutipes

Kotake

Hirata

Degi

et al. 2011

Journal of Biological Chemistry

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Arabinogalactan proteins are proteoglycans found on the cell surface and in the cell walls of higher plants. The carbohydrate moieties of most arabinogalactan proteins are composed of ␤-1,3-galactan main chains and ␤-1,6-galactan side chains, to which other auxiliary sugars are attached. For the present study, an endo-␤-1,3-galactanase, designated FvEn3GAL, was first purified and cloned from winter mushroom Flammulina velutipes. The enzyme specifically hydrolyzed ␤-1,3-galactan, but did not act on ␤-1,3-glucan, ␤-1,3:1,4-glucan, xyloglucan, and agarose. It released various ␤-1,3-galactooligosaccharides together with Gal from ␤-1,3-galactohexaose in the early phase of the reaction, demonstrating that it acts on ␤-1,3-galactan in an endo-fashion. Phylogenetic analysis revealed that FvEn3GAL is member of a novel subgroup distinct from known glycoside hydrolases such as endo-␤-1,3-glucanase and endo-␤-1,3:1,4-glucanase in glycoside hydrolase family 16. Point mutations replacing the putative catalytic Glu residues conserved for enzymes in this family with Asp abolished activity. These results indicate that FvEn3GAL is a highly specific glycoside hydrolase 16 endo-␤-1,3-galactanase.

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“…A mixture of iso-maltooligosaccharides were prepared from dextran by mild acid hydrolysis (Nishigaki et al, 1978) and derivatized with ABEE as described previously (Matsuura and Imaoka, 1988). The structures of Asn-linked oligosaccharides uscd in this study are listed in Table 1.…”

Section: Introductionmentioning

confidence: 99%

“…Oligosaccharides, liberated from these glycoproteins by hydrazinolysis and N-reacetylation. were labelled with ABEE as described previously (Matsuura and Imaoka, 1988). The ABEEsialyloligosaccharides obtained from cach glycoprotein were desialylated by the treatment with A .…”

Section: Introductionmentioning

confidence: 99%

The combination of normal phase with reversed phase high performance liquid chromatography for the analysis of asparagine‐linked neutral oligosaccharides labelled with p‐aminobenzoic ethyl ester

Matsuura

Ohta

Murakami

et al. 1992

Biomedical Chromatography

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We have developed a novel approach for the analysis of asparagine-linked neutral oligosaccharides derived from glycoproteins. The oligosaccharides are labelled with p-aminobenzoic ethyl ester and the derivatives are separated on two high performance liquid chromatographic columns, one containing amide-silica and the other containing octadecyl-silica. The elution positions of 39 different ABEE-oligosaccharides on the two columns were plotted on a two-dimensional map. Unique non-overlapping positions of these oligosaccharides demonstrate that this technology would be useful for the identification of Asn-linked oligosaccharides at high sensitivity.

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Chromatographic separation of asparagine-linked oligosaccharides labeled with an ultravioletabsorbing compound,p-aminobenzoic acid ethyl ester

Cited by 43 publications

References 27 publications

β-Galactosyl Yariv Reagent Binds to the β-1,3-Galactan of Arabinogalactan Proteins

β-Galactosyl Yariv Reagent Binds to the β-1,3-Galactan of Arabinogalactan Proteins

Endo-β-1,3-galactanase from Winter Mushroom Flammulina velutipes

The combination of normal phase with reversed phase high performance liquid chromatography for the analysis of asparagine‐linked neutral oligosaccharides labelled with p‐aminobenzoic ethyl ester

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