Comparative analysis of oligosaccharide specificities of fucose-specific lectins from Aspergillus oryzae and Aleuria aurantia using frontal affinity chromatography.
Aleuria aurantia lectin (AAL) is widely used to estimate the extent of alpha1,6-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 alpha1,6-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 (K(d)) 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 alpha1,6-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 (K(a)) for alpha1,6-fucosylated oligosaccharides than AAL and only AAL additionally recognized oligosaccharides which were alpha1,3-fucosylated at the reducing terminal GlcNAc. These results explain why AOL reflects the extent of alpha1,6-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.
Matsumura
Kengo
K
Research Institute, Gekkeikan Sake Co. Ltd., 300 Katahara-cho, Fushimi-ku, Kyoto 612-8361, Japan. kengo@gekkeikan.co.jp
Higashida
Katsuya
K
Hata
Yoji
Y
Kominami
Junko
J
Nakamura-Tsuruta
Sachiko
S
Hirabayashi
Jun
J
D001203:Ascomycota / Q000378:metabolism*; D001236:Aspergillus oryzae / Q000378:metabolism*; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods*; D007700:Kinetics; D037102:Lectins / Q000737:chemistry* / Q000378:metabolism; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity
D037102:Lectins; D009844:Oligosaccharides; C064549:lectin, Aleuria aurantia