Yuen C T, Lawson A M, Chai W, Larkin M, Stoll M S, Stuart A C, Sullivan F X, Ahern T J, Feizi T
Glycoconjugates Section, MRC Clinical Research Centre, Harrow, Middlesex, U.K.
Biochemistry. 1992 Sep 29;31(38):9126-31. doi: 10.1021/bi00153a003.
E-selectin is the inducible adhesion protein on the surface of endothelial cells which has a crucial role in the initial stages of recruitment of leucocytes to sites of inflammation. In addition, it is almost certainly involved in tumor cell adhesion and metastasis. This report is concerned with identification of a new class of oligosaccharide ligand--sulfate-containing--for the human E-selectin molecule from among oligosaccharides on an ovarian cystadenoma glycoprotein. This has been achieved by application of the neoglycolipid technology to oligosaccharides released from the glycoprotein by mild alkaline beta-elimination. Oligosaccharides were conjugated to lipid, resolved by thin-layer chromatography, and tested for binding by Chinese hamster ovary cells which had been transfected to express the full-length E-selectin molecule. Several components with strong E-selectin binding activity were revealed among acidic oligosaccharides. The smallest among these was identified by liquid secondary ion mass spectrometric analysis of the neoglycolipid, in conjunction with methylation analysis of the purified oligosaccharide preparation as an equimolar mixture of the Le(a)- and Le(x)/SSEA-1-type fucotetrasaccharides sulfated at position 3 of outer galactose: [formula: see text] To our knowledge this is the first report of a sulfofucooligosaccharide ligand for E-selectin. The binding activity is substantially greater than those of lipid-linked Le(a) and Le(x)/SSEA-1 sequences and is at least equal to that of the 3'-sialyl-Le(x)/SSEA-1 glycolipid analogue.
E-选择素是内皮细胞表面的诱导性黏附蛋白，在白细胞募集到炎症部位的初始阶段起着关键作用。此外，它几乎肯定参与肿瘤细胞的黏附和转移。本报告关注从卵巢囊腺瘤糖蛋白的寡糖中鉴定出一类新的人E-选择素分子含硫酸盐的寡糖配体。这是通过将新糖脂技术应用于经温和碱性β-消除从糖蛋白释放的寡糖来实现的。寡糖与脂质偶联，通过薄层色谱分离，并通过转染以表达全长E-选择素分子的中国仓鼠卵巢细胞进行结合测试。在酸性寡糖中发现了几种具有强E-选择素结合活性的成分。其中最小的通过新糖脂的液相二次离子质谱分析以及纯化寡糖制剂的甲基化分析鉴定为在外半乳糖3位硫酸化的Le(a)-和Le(x)/SSEA-1型岩藻四糖的等摩尔混合物：[化学式：见正文]据我们所知，这是关于E-选择素的硫酸化岩藻寡糖配体的首次报道。其结合活性远大于脂质连接的Le(a)和Le(x)/SSEA-1序列，且至少与3'-唾液酸-Le(x)/SSEA-1糖脂类似物相当。
