glycans of soybean peroxidase

• Published in: Glycobiology (Oxford) 1996-01, Vol.6 (1), p.23-32
• Main Authors: Gray, J.S.S, Yang, B.Y, Hull, S.R, Venzke, D.P, Montgomery, R
• Format: Article
• Language: English
• Subjects: Amidohydrolases - metabolism; Carbohydrate Sequence; chemical analysis; Chromatography, Ion Exchange; glycan; Glycine max - enzymology; Glycoproteins - chemistry; Glycoproteins - metabolism; hulls; Hydrogen-Ion Concentration; MALDI-TOF; Methylation; methylation analysis; Molecular Sequence Data; Molecular Weight; N-linked oligosaccharides; oligosaccharides; Oligosaccharides - analysis; Oligosaccharides - chemistry; Oligosaccharides - metabolism; Peptide Mapping; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; Peroxidase - analysis; Peroxidase - chemistry; Peroxidase - metabolism; peroxidases; Polysaccharides - analysis; Polysaccharides - chemistry; soybean peroxidase; soybeans; Trypsin - metabolism
• ISSN: 0959-6658; 1460-2423
• DOI: 10.1093/glycob/6.1.23

Soybean hull peroxidase (SBP, E.C. 1.11.1.7), an anionic glycoprotein, was found to contain 18.2% carbohydrate with the average composition: 2 mol GlcNAc, 3.3 mol Man, 0.9 mol Fuc, and 0.7 mol Xyl. The oligosaccharides of SBP, after release with glycopeptidase A, were investigated by a combination of high pH anion exchange chromatography with pulsed amperometric detection, methylation analysis and matrix assisted laser desorption/ionization-time-of-flight mass spectrometry. The structure of the major oligo-saccharide, accounting for 60 to 65% of the total, is Man α1 → 6(Manβ1 → 3)(Xylβ1 → 2)Manβ1 →4GlcNAcβ1 → 4(Fucα1 → 3)GlcNAc. A further 20 to 25% of the released oligosaccharides belong to the (Xyl)x Manm(Fuc)rGlcNAc2 (m = 2, 4, 5, 6; f = 0 or 1; x = 0 or 1) family. The rest of the oligosaccharides were of the high-mannose type. Investigation of the six tryptic fractions containing carbohydrate revealed considerable heterogeneity in the N-linked oligosaccharides present in each fraction. The major glycan (4,Table III) was present in each fraction. Two of the fractions contained the major part of the high-mannose type glycans, Man7GlcNAc2 (m = 5–9), the major species being Man7GlcNAc2. The other four fractions contained mainly members of the (Xyl)xManm(Fuc)rGlcNAc2 (m = 2, 4, 5, 6; f = 0 or 1; x = 0 or 1) family. Methylation analysis of the holo- and apo-SBP provide support for the structures proposed for the oligosaccharides as well as for the heterogeneity of the glycopeptide fractions.