Osanai T, Feizi T, Chai W, Lawson A M, Gustavsson M L, Sudo K, Araki M, Araki K, Yuen C T
Glycosciences Laboratory, Northwick Park Hospital, Harrow, Middlesex, UK.
Biochem Biophys Res Commun. 1996 Jan 17;218(2):610-5. doi: 10.1006/bbrc.1996.0108.
In search of endogenous oligosaccharide ligands for the endothelial adhesion molecule E-selectin in mouse, glycolipids from tissues and the neutrophilic cell line 32D c13 were tested for E-selectin binding. Kidneys of BALB/c and NMRI mice (but not CBA) and the 32D c13 cells were found to contain minor glycolipid populations that support strongly the binding of murine E-selectin. By chromatogram binding experiments and in situ liquid secondary ion mass spectrometry (LSIMS) with neoglycolipids derived from their endoglycoceramidase-released oligosaccharides, in conjunction with compositional and linkage analyses, one of the glycolipid ligands in kidney was identified as the Le(x)-active extended globo-glycolipid: [formula: see text] Neoglycolipids enriched for the ligand structures were obtained from oligosaccharides released by endo-beta-galactosidase from the 32D c13 cells. By TLC-LSIMS and antibody binding, the main E-selectin binding determinant on these was identified as sialyl-Le(a).
为了寻找小鼠体内内皮细胞粘附分子E-选择素的内源性寡糖配体，对来自组织和嗜中性细胞系32D c13的糖脂进行了E-选择素结合测试。发现BALB/c和NMRI小鼠（但不是CBA小鼠）的肾脏以及32D c13细胞含有少量糖脂群体，这些糖脂能强烈支持鼠源E-选择素的结合。通过色谱结合实验以及对其内切糖神经酰胺酶释放的寡糖衍生的新糖脂进行原位液体二次离子质谱分析（LSIMS），结合组成和连接分析，确定肾脏中的一种糖脂配体为具有Le(x)活性的延伸球型糖脂：[分子式：见正文] 从32D c13细胞经β-半乳糖苷内切酶释放的寡糖中获得了富含配体结构的新糖脂。通过薄层层析-液体二次离子质谱分析（TLC-LSIMS）和抗体结合，确定这些新糖脂上主要的E-选择素结合决定簇为唾液酸化-Le(a)。
