Kushi Y, Arita M, Ishizuka I, Kasama T, Fredman P, Handa S
Department of Biochemistry, Faculty of Medicine, Tokyo Medical and Dental University, Japan.
Biochim Biophys Acta. 1996 Dec 13;1304(3):254-62. doi: 10.1016/s0005-2760(96)00125-7.
A novel sulfated glycosphingolipid containing a sulfated galactosyl residue was isolated from bovine erythrocyte ghosts, and purified to homogeneity by column chromatography on DEAE-Sephadex and silica beads. Structural characterization included compositional analyses, permethylation studies, proton nuclear magnetic resonance (NMR) spectroscopy, negative secondary ion mass spectrometry (SIMS), solvolysis and immunostaining on thin-layer chromatogram. As a result, the structure of this glycolipid is proposed as HSO3-Gal beta 1-1 Cer. The ceramide portion contained d18:1, d18:0 and t18:0, and the predominant fatty acid consisted of palmitate and palmitate with a hydroxy group, as deduced by both compositional analysis and negative SIMS mass spectrometry. The component of this glycosphingolipid probably originates from erythrocytes and platelets as indicated by the results of flow cytometry analysis using Sulph I monoclonal antibody. The yield of galactosyl sulfatide was about 0.37 mg/kg wet bovine erythrocyte membranes, about three times that of human kidney. Our results strongly suggest that galactosylceramide sulfate on erythroid cells may play an important biological role in cell to cell interaction and recognition.
从牛红细胞膜中分离出一种含有硫酸化半乳糖基残基的新型硫酸化糖鞘脂,并通过DEAE-葡聚糖凝胶柱色谱和硅胶柱色谱将其纯化至均一。结构表征包括成分分析、全甲基化研究、质子核磁共振(NMR)光谱、负离子二次离子质谱(SIMS)、溶剂解以及薄层色谱上的免疫染色。结果表明,这种糖脂的结构被确定为HSO3-Gal β1-1 Cer。通过成分分析和负离子SIMS质谱推断,神经酰胺部分包含d18:1、d18:0和t18:0,主要脂肪酸由棕榈酸和带有羟基的棕榈酸组成。使用Sulph I单克隆抗体进行的流式细胞术分析结果表明,这种糖鞘脂的成分可能源自红细胞和血小板。半乳糖基硫酸脑苷脂的产量约为0.37 mg/kg湿牛红细胞膜,约为人肾产量的三倍。我们的结果强烈表明,红细胞上的硫酸化半乳糖神经酰胺可能在细胞间相互作用和识别中发挥重要的生物学作用。