Müthing J, Spanbroek R, Peter-Katalinić J, Hanisch F G, Hanski C, Hasegawa A, Unland F, Lehmann J, Tschesche H, Egge H
Institute of Cell Culture Technology, University of Bielefeld, Germany.
Glycobiology. 1996 Mar;6(2):147-56. doi: 10.1093/glycob/6.2.147.
The isolation and structural characterization of fucosylated neolacto-series gangliosides with linear poly-N-acetyllactosaminyl chains from normal human granulocytes is described. Gangliosides were purified by consecutive use of anion exchange HPLC on Fractogel TMAE-650(S), adsorption and reversed phase HPLC on Nucleosil 50-7 and Nucleosil 7C18 columns, respectively. TLC immunostaining with carbohydrate specific monoclonal antibodies, fast atom bombardment-mass spectrometry (FAB-MS) of the permethylated derivatives and gas chromatography-electron impact mass spectrometry (GC-EIMS) of partially methylated alditol acetates were used for structure elucidations. One ganglioside was identified as sialyl Lewis(x) antigen with nLcOse6Cer core, Neu5-Ac alpha 2-3Gal beta 1-4 (Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc NAc beta 1-3 Gal beta 1-4Glc beta 1-1Cer. Furthermore, monosialylated ceramide deca-, undeca-, dodeca- and tridecasaccharides with three (nLcOse8Cer) and four (nLcOse10Cer) linear lactosaminyl repeats were identified, carring one to three fucoses. The ceramide portions were found to contain C18 sphingosine and predominantly C16:0 fatty acids. All monosialogangliosides were homogenous concerning their terminal alpha 2-3 Neu5Ac-sialylation, but different in their fucosylation status. Beside VI3Neu5Ac, V3Fuc-nLcOse6Cer, in two of the fucosylated polylactosaminyl ganglioside fractions the sialyl Lewis(x) epitope was found, whereas five species expressed the terminal VIM-2 motif. The role of protein linked sialy Lewis(x) epitope of human granulocytes as a ligand for endothelial leukocyte adhesion molecule-1 (ELAM-1; E-selectin) and platelet activation-dependent granule external membrane protein (PADGEM; P-selectin) is well documented. However, the involvement of endothelial cells E-and/or P-selectin mediated cell-cell adhesion via lipid bound sialyl Lewis(x) and/or VIM-2 epitopes on human granulocytes has to be proved in further investigations.
本文描述了从正常人粒细胞中分离和鉴定具有线性聚-N-乙酰乳糖胺链的岩藻糖基化新乳糖系列神经节苷脂及其结构特征。神经节苷脂的纯化分别通过在Fractogel TMAE-650(S)上连续使用阴离子交换高效液相色谱（HPLC）、在Nucleosil 50-7和Nucleosil 7C18柱上进行吸附和反相HPLC来实现。使用碳水化合物特异性单克隆抗体进行薄层层析免疫染色、对全甲基化衍生物进行快原子轰击质谱（FAB-MS）以及对部分甲基化糖醇乙酸酯进行气相色谱-电子轰击质谱（GC-EIMS）来阐明结构。一种神经节苷脂被鉴定为具有nLcOse6Cer核心的唾液酸化路易斯（x）抗原，即Neu5-Acα2-3Galβ1-4（Fucα1-3）GlcNAcβ1-3Galβ1-4Glc NAcβ1-3 Galβ1-4Glcβ1-1Cer。此外，还鉴定出了具有三个（nLcOse8Cer）和四个（nLcOse10Cer）线性乳糖胺重复序列且带有一到三个岩藻糖的单唾液酸化神经酰胺十糖、十一糖、十二糖和十三糖。发现神经酰胺部分含有C18鞘氨醇且主要为C16:0脂肪酸。所有单唾液酸神经节苷脂在其末端α2-3 Neu5Ac-唾液酸化方面是均一的，但在岩藻糖基化状态上有所不同。除了VI3Neu5Ac、V3Fuc-nLcOse6Cer外，在两个岩藻糖基化的多乳糖胺神经节苷脂组分中发现了唾液酸化路易斯（x）表位，而有五个种类表达了末端VIM-2基序。人粒细胞中与蛋白质相连的唾液酸化路易斯（x）表位作为内皮细胞白细胞粘附分子-1（ELAM-1；E-选择素）和血小板活化依赖性颗粒外膜蛋白（PADGEM；P-选择素）的配体的作用已得到充分证明。然而内皮细胞通过人粒细胞上脂质结合的唾液酸化路易斯（x）和/或VIM-2表位介导的细胞间粘附的参与情况还有待进一步研究证实。
