Comparative analysis of oligosaccharide specificities of fucose-specific lectins from Aspergillus oryzae
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- 作者：Kengo Matsumura ; Katsuya Higashida ; Yoji Hata ; Junko Kominami ; Sachiko Nakamura-Tsuruta ; Jun Hirabayashi
- 关键词：Oligosaccharide specificity ; Frontal affinity chromatography ; α ; 1 ; 6-Fucosylation ; Lectin ; Multivalent carbohydrate recognition
- 刊名：Analytical Biochemistry
- 出版年：2009
- 出版时间：15 March 2009
- 年：2009
- 卷：386
- 期：2
- 页码：217-221
- 全文大小：224 K
文摘
Aleuria aurantia lectin (AAL) is widely used to estimate the extent of α1,6-fucosylated oligosaccharides and to fractionate glycoproteins for the detection of specific biomarkers for developmental antigens. Our previous studies have shown that Aspergillus oryzae lectin (AOL) reflects the extent of α1,6-fucosylation more clearly than AAL. However, the subtle specificities of these lectins to fucose linked to oligosaccharides through the 2-, 3-, 4-, or 6-position remain unclear, because large amounts of oligosaccharides are required for the systematic comparative analysis using surface plasmon resonance. Here we show a direct comparison of the dissociation constants (Kd) of AOL and AAL using 113 pyridylaminated oligosaccharides with frontal affinity chromatography. As a result, AOL showed a similar specificity as AAL in terms of the high affinity for α1,6-fucosylated oligosaccharides, for smaller fucosylated oligosaccharides, and for oligosaccharides fucosylated at the reducing terminal core GlcNAc. On the other hand, AOL showed 2.9–6.2 times higher affinity constants (Ka) for α1,6-fucosylated oligosaccharides than AAL and only AAL additionally recognized oligosaccharides which were α1,3-fucosylated at the reducing terminal GlcNAc. These results explain why AOL reflects the extent of α1,6-fucosylation on glycoproteins more clearly than AAL. This systematic comparative analysis made from a quantitative viewpoint enabled a clear physical interpretation of these fucose-specific lectins with multivalent fucose-binding sites.