title: Carbohydrate binding specificity of a fucose-specific lectin from Aspergillus oryzae: a novel probe for core fucose.
abstract: The alpha1,6-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 alpha1,6-fucosylated chain among alpha1,2-, alpha1,3-, alpha1,4-, and alpha1,6-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 alpha1,2-fucosylated PA-sugar chain (H antigen) relative to the alpha1,6-fucosylated chain was weaker than that of AAL, and that the interactions of AOL and AAL with alpha1,6-fucosylated glycopeptide (FGP), which is considered more similar to in vivo glycoproteins than PA-sugar chains, were similar to their interactions with the alpha1,6-fucosylated PA-sugar chain. Furthermore, positive staining of AOL, but not AAL, was completely abolished in the cultured embryo fibroblast (MEF) cells obtained from alpha1,6-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.
authors: Research Institute, Gekkeikan Sake Company Ltd., 300 Katahara-cho, Fushimi-ku, Kyoto 612-8361, Japan. kengo@gekkeikan.co.jp
authors: Kengo
authors: K
authors: Matsumura
authors: Katsuya
authors: K
authors: Higashida
authors: Hiroki
authors: H
authors: Ishida
authors: Yoji
authors: Y
authors: Hata
authors: Kenji
authors: K
authors: Yamamoto
authors: Masaki
authors: M
authors: Shigeta
authors: Yoko
authors: Y
authors: Mizuno-Horikawa
authors: Xiangchun
authors: X
authors: Wang
authors: Eiji
authors: E
authors: Miyoshi
authors: Jianguo
authors: J
authors: Gu
authors: Naoyuki
authors: N
authors: Taniguchi
mesh_terms: D000818:Animals; D001236:Aspergillus oryzae / Q000737:chemistry*; D004622:Embryo, Mammalian / Q000166:cytology / Q000201:enzymology; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D005643:Fucose / Q000737:chemistry* / Q000378:metabolism; D005647:Fucosyltransferases / Q000172:deficiency; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D037102:Lectins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding / Q000502:physiology; D013379:Substrate Specificity / Q000502:physiology
chemical_list: D005656:Fungal Proteins; D006023:Glycoproteins; D037102:Lectins; D015335:Molecular Probes; D005643:Fucose; D005647:Fucosyltransferases; C020559:Glycoprotein 6-alpha-L-fucosyltransferase