Nagatsuka Y, Kasama T, Ohashi Y, Uzawa J, Ono Y, Shimizu K, Hirabayashi Y
Department of Immunology and Microbiology, Nihon University School of Medicine, Tokyo, Japan.
FEBS Lett. 2001 May 25;497(2-3):141-7. doi: 10.1016/s0014-5793(01)02464-4.
Cord red cell membranes express many differentiation-related molecules. To study such molecules, we have established human cell lines, termed GL-1 and GL-2, by the Epstein-Barr virus transformation method, both of which produce monoclonal anti-i cold agglutinin [Y. Nagatsuka et al., Immunol. Lett. 46 (1995) 93-100]. Thin layer chromatography immunoblotting analysis revealed that these antibodies had broad specificities reacting with a variety of glycolipid antigens. Of the immunoreactive lipid antigens, a new phosphoglycerolipid containing glucose from human cord red cells was found. The isolated lipid was unstable to alkaline hydrolysis and contained glucose as a sole sugar. Secondary ion mass spectrum-collision-induced dissociation mass spectrometric analysis of this lipid gave the main molecular ion peak at m/z 885 corresponding to phosphatidylhexose. This antigen was susceptible to phospholipases A2, C and D but resistant to phosphatidylinositol-specific phospholipase C. Two-dimensional nuclear magnetic resonance spectroscopy confirmed that glucose is linked to the sn-glycerol 3-phosphate residue with a beta-anomeric configuration. Based upon these combined results, we identified this lipid as phosphatidyl-beta-D-glucose. This is the first report showing the presence of the glucosylated glycerophospholipid in mammalian sources.
脐带红细胞膜表达许多与分化相关的分子。为了研究这些分子,我们通过爱泼斯坦-巴尔病毒转化法建立了人细胞系,命名为GL-1和GL-2,这两种细胞系都产生单克隆抗-i冷凝集素[Y. Nagatsuka等人,《免疫学快报》46 (1995) 93-100]。薄层色谱免疫印迹分析表明,这些抗体具有广泛的特异性,能与多种糖脂抗原发生反应。在免疫反应性脂质抗原中,发现了一种来自人脐带红细胞的含葡萄糖的新型磷酸甘油脂。分离出的脂质对碱性水解不稳定,且仅含葡萄糖作为唯一的糖。对该脂质进行二次离子质谱-碰撞诱导解离质谱分析,得到对应于磷脂酰己糖的主要分子离子峰,质荷比为885。该抗原对磷脂酶A2、C和D敏感,但对磷脂酰肌醇特异性磷脂酶C有抗性。二维核磁共振光谱证实葡萄糖以β-异头构型与sn-甘油3-磷酸残基相连。基于这些综合结果,我们将这种脂质鉴定为磷脂酰-β-D-葡萄糖。这是首次报道在哺乳动物来源中存在糖基化甘油磷脂。