Suzuki Y, Toda Y, Tamatani T, Watanabe T, Suzuki T, Nakao T, Murase K, Kiso M, Hasegawa A, Tadano-Aritomi K
Department of Biochemistry, University of Shizuoka School of Pharmaceutical Science, Japan.
Biochem Biophys Res Commun. 1993 Jan 29;190(2):426-34. doi: 10.1006/bbrc.1993.1065.
Specific sugar ligands which bind to rat L-selectin (LECAM-1, lymphocyte homing receptor)-IgG chimera were investigated by thin-layer chromatography-binding assay and ELISA. Rat L-selectin-IgG bound to native sulfated glycolipids such as sulfatide (I3SO3-GalCer), seminolipid, SM3(II3SO3-LacCer), SB2(III3SO3,II3SO3-Gg3Cer), SB1a (IV3SO3,II3SO3-Gg4Cer). The binding activity of the SM2 (II3SO3-Gg3Cer) carrying the internal sulfated Gal was very low, indicating that non-reducing terminal SO3H-3Gal beta 1- structure is essential for the binding. The reactivity of sulfatide was higher than that of sialyl Le(x) [Neu5Ac alpha 2-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc beta 1-Ceramide] either in the presence of 1 mM Ca2+ or 1 mM Ca2+ and 5 mM EGTA. All the non-fucosylated gangliosides including ganglio-series and lacto-series type I and II chains tested gave a negative reaction. Neutral glycosphingolipids tested were all negative. Binding assay using synthetic sulfatide analogs indicated that the L-selectin-binding to its sulfated sugar ligands is position specific and depends on the number of the sulfate group, i.e., (SO3)3-3,4,6Gal- > (SO3)2-3,6Gal- > SO3-3Gal- > SO3-3Glc- > SO3-6Gal- > SO3-2Gal-. Lower reactivity of the chimera to SO3-3Glc-rather than SO3-3Gal- indicates that the chimera recognizes the configuration of hydroxyl group linked to the 4 carbon atom of the pyranose ring in galactose.
通过薄层色谱结合试验和酶联免疫吸附测定法研究了与大鼠L-选择素（LECAM-1，淋巴细胞归巢受体）-IgG嵌合体结合的特定糖配体。大鼠L-选择素-IgG与天然硫酸化糖脂结合，如硫苷脂（I3SO3-GalCer）、半乳糖神经酰胺、SM3（II3SO3-LacCer）、SB2（III3SO3，II3SO3-Gg3Cer）、SB1a（IV3SO3，II3SO3-Gg4Cer）。带有内部硫酸化半乳糖的SM2（II3SO3-Gg3Cer）的结合活性非常低，这表明非还原末端SO3H-3Galβ1-结构对于结合至关重要。在存在1 mM Ca2+或1 mM Ca2+和5 mM乙二醇双（2-氨基乙基醚）四乙酸（EGTA）的情况下，硫苷脂的反应性均高于唾液酸化Le(x) [Neu5Acα2-3Galβ1-4(Fucα1-3)GlcNAcβ1-3Galβ1-4Glcβ1-神经酰胺]。所测试的所有非岩藻糖基化神经节苷脂，包括神经节系列和乳糖系列I型和II型链，均呈阴性反应。所测试的中性糖鞘脂均为阴性。使用合成硫苷脂类似物的结合试验表明，L-选择素与其硫酸化糖配体的结合具有位置特异性，并取决于硫酸根基团的数量，即(SO3)3-3,4,6Gal- > (SO3)2-3,6Gal- > SO3-3Gal- > SO3-3Glc- > SO3-6Gal- > SO3-2Gal-。嵌合体对SO3-3Glc-的反应性低于对SO3-3Gal-的反应性，这表明嵌合体识别与吡喃糖环4碳原子相连的羟基的构型在半乳糖中。
