Naiki M, Fong J, Ledeen R, Marcus D M
Biochemistry. 1975 Nov 4;14(22):4831-7. doi: 10.1021/bi00693a009.
A glycosphingolipid with blood group P1 activity was extracted from an acetone powder of human erythrocyte stroma with chloroform-methanol. It was purified by chromatography on columns of silicic acid and by preparative thin-layer chromatography of the fully acetylated and deacetylated glycolipid. The purified glycolipid contained galactose, N-acetylglucosamine, and glucose in a molar ratio of 3:1:1. Treatment of the P1 glycolipid with fig alpha-galactosidase released a single galactosyl residue and destroyed the blood group activity, and the alpha-galactosidase product had the same chromatographic mobility as paragloboside. Substitution sites on the neutral sugars of the P1 glycolipid and the alpha-galactosidase product were established by identification of methylated alditol acetates, and substitution on N-acetylglucosamine was determined by identification of methyl glycoside derivatives. The terminal nonreducing disaccharide of the P1 glycolipid is Gal(alpha, 1 leads to 4)Gal. N-Acetylglucosamine was identified as the next sugar in sequence by mass spectrometric analysis of the permethylated P1 glycolipid. On the assumption that the glucose residue is linked to ceramide, we propose the following structure for the P1 glycolipid: Gal(alpha, 1 leads to 4)Gal(beta, 1 leads to 4)Glc-NAc(beta, 1 leads to 4)Glc-Cer.
用氯仿 - 甲醇从人红细胞基质的丙酮粉中提取了具有血型P1活性的糖鞘脂。通过硅胶柱色谱以及对完全乙酰化和脱乙酰化糖脂进行制备型薄层色谱对其进行纯化。纯化后的糖脂中半乳糖、N - 乙酰葡糖胺和葡萄糖的摩尔比为3:1:1。用无花果α - 半乳糖苷酶处理P1糖脂会释放出一个单一的半乳糖基残基并破坏血型活性，且α - 半乳糖苷酶产物与副球蛋白的色谱迁移率相同。通过鉴定甲基化糖醇乙酸酯确定了P1糖脂和α - 半乳糖苷酶产物中性糖上的取代位点，通过鉴定甲基糖苷衍生物确定了N - 乙酰葡糖胺上的取代情况。P1糖脂的末端非还原二糖为Gal(α,1→4)Gal。通过对全甲基化P1糖脂的质谱分析确定N - 乙酰葡糖胺为序列中的下一个糖。假设葡萄糖残基与神经酰胺相连，我们提出P1糖脂的如下结构：Gal(α,1→4)Gal(β,1→4)Glc - NAc(β,1→4)Glc - Cer。
