Carbohydrate Binding Specificity of a Fucose-specific Lectin from Aspergillus oryzae

Abstract

The ␣1,6-fucosyl residue (core fucose) of glycoproteins is widely distributed in mammalian tissues and is altered under pathological conditions. A probe that specifically detects core fucose is important for understanding the role of this oligosaccharide structure. Aleuria aurantia lectin (AAL) and Lens culimaris agglutinin-A (LCA) have been often used as carbohydrate probes for core fucose in glycoproteins. Here we show, by using surface plasmon resonance (SPR) analysis, that Aspergillus oryzae L-fucose-specific lectin (AOL) has strongest preference for the ␣1,6-fucosylated chain among ␣1,2-, ␣1,3-, ␣1,4-, and ␣1,6-fucosylated pyridylaminated (PA)-sugar chains. These results suggest that AOL is a novel probe for detecting core fucose in glycoproteins on the surface of animal cells. A comparison of the carbohydrate-binding specificity of AOL, AAL, and LCA by SPR showed that the irreversible binding of AOL to the ␣1,2-fucosylated PA-sugar chain (H antigen) relative to the ␣1,6-fucosylated chain was weaker than that of AAL, and that the interactions of AOL and AAL with ␣1,6-fucosylated glycopeptide (FGP), which is considered more similar to in vivo glycoproteins than PA-sugar chains, were similar to their interactions with the ␣1,6-fucosylated PA-sugar chain. Furthermore, positive staining of AOL, but not AAL, was completely abolished in the cultured embryo fibroblast (MEF) cells obtained from ␣1,6-fucosyltransferase (Fut8) knock-out mice, as assessed by cytological staining. Taken together, these results suggest that AOL is more suitable for detecting core fucose than AAL or LCA.

A Novel Core Fucose-specific Lectin from the Mushroom Pholiota squarrosa

“…␣1-6 fucosylation is one of the most important oligosaccharide modifications in carcinogenesis; however, although many studies related to fucosylation have been conducted, they have not completely clarified the difference between ␣1-2, ␣1-3, or ␣1-4 fucosylation and ␣1-6 fucosylation. A hindrance to this clarification has been the lack of a tool for the specific detection of ␣1-6 fucosyl linkage; AAL, which is used in many studies, recognizes all types of fucosyl linkages (13,17).…”

“…LCA can be used for affinity chromatography, but using it for lectin blot analysis to evaluate cellular fucosylation can be difficult because of its low sugar binding specificity. Conventionally, in addition to LCA, other commercially available core fucose-binding lectins, such as Pisum sativum agglutinin (9), Aleuria aurantia lectin (AAL) (10 -13), Narcissus pseudonarcissus agglutinin, Vicia faba agglutinin (14 -16), and Aspergillus oryzae lectin (12,17,18) have been used in studies on glycobiology. However, most fucose-binding lectins recognize any type of fucosylation, and LCA binds not only to fucose but also to mannose residues in N-glycans (3).…”

Optimization of culture conditions and characterization of a new lectin from Aspergillus niger

“…Fungal lectins have been found to be specifi c for mucin and fetuin [12,13]. Among the genus Aspergillus, lectins have been reported from A. fumigatus [14,15] and A. oryzae [16]. In the present study, we investigated a new mucin-specifi c lectin from the mycelium of A. niger.…”

Antimicrobial activity of lectins from Antimicrobial Activity of Lectins from Plants

“…For research a novel probe for core fucose from Aspergillus oryzae L-fucose-specific lectin (AOL) has strongest preference for the alpha 1,6-fucosylated chain among α-1,2-, α-1,3-, α-1,4-, and α-1,6-fucosylated pyridylaminated (PA)-sugar chains. These results suggest that AOL is a novel probe for detecting core fucose in glycoproteins on the surface of animal cells (Matsumura et al, 2007). Furthermore, Lotus tetragonolobus lectin is a fucose-specific legume lectin.…”