Abstract
The structural requirements for the interaction of asparagine-linked oligosaccharide moieties of glycoproteins withErythrina variegata agglutinin (EVA) were investigated by means of affinity chromatography on an EVA-Sepharose column. Some of the branched poly-N-acetyllactosamine-type oligosaccharides obtained from human erythrocyte band 3 glycoprotein were found to show high affinity to EVA-Sepharose, whereas complex-type oligosaccharides were shown to have low affinity. Hybrid type, oligomannose-type and unbranched poly-N-acetyllactosamine-type oligosaccharides bound very little or not at all to EVA-Sepharose. To further study the carbohydrate-binding specificity of this lectin, we investigated the interaction of immobilized EVA and oligosaccharide fragments obtained through partial hydrolysis from branched poly-N-acetyllactosamine-type oligosaccharides. Branched poly-N-acetyllactosamine-type oligosaccharides were subjected to limited hydrolysis with 0.1% trifluoroacetic acid at 100°C for 40 min and then separated on an amino-bonded silica column. One of pentasaccharides thus prepared strongly bound to the EVA-Sepharose column. Structural analysis of this pentasaccharide showed that the Galβ1-4GlcNAcβ1-3(Galβ1-4GlcNAcβ1-6)Gal sugar sequence, which is an l-antigen determinant, was essential for the high affinity binding of the oligosaccharides to the EVA-Sepharose column.
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Abbreviations
- EVA:
-
Erythrina variegata agglutinin
- WGA:
-
wheat germ agglutinin
- STA:
-
potato lectin
- LEA:
-
tomato lectin
- DSA:
-
Datura stramonium agglutinin
- PBS:
-
0.01 M sodium phosphate buffer, pH 7.3, containing 0.15 M NaCl
- Galol:
-
galactitol
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Li, H., Yamamoto, K., Kawashima, H. et al. Structural requirements for the binding of oligosaccharides to immobilized lectin ofErythrina variegata (Linn) var.Orientalis . Glycoconjugate J 7, 311–322 (1990). https://doi.org/10.1007/BF01073375
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DOI: https://doi.org/10.1007/BF01073375