Xu X, Stack R J, Rao N, Caulfield J P
Syntex Discovery Research, Palo Alto, California 94304.
Exp Parasitol. 1994 Nov;79(3):399-409. doi: 10.1006/expr.1994.1102.
The glycocalyx (GCX) that covers schistosomal cercariae is a complex molecule that has immunomodulating properties. Here, we purified milligram amounts of GCX using Anguilla lectin which binds to the GCX covering the cercarial body and tail. Typically, 10 million cercariae were extracted with phenol, dialyzed, and chromatographed on a Sepharose 2B-CL column. An average of 39 mg of total carbohydrate eluted near the void volume from which 31 mg of glycogen-like material was further separated by lectin affinity chromatography. Its identity was established by compositional analysis, sensitivity to amylase digestion, and its nuclear magnetic resonance spectrum. The lectin-bound GCX was eluted with 0.1 M fucose with a final yield of 5.3 mg carbohydrate. Fucose composed 40% of the total GCX carbohydrate with lesser but approximately equal amounts of galactose, glucosamine, and galactosamine present. NMR data indicated that the amino sugars were N-acetylated. Glucose was also present but in varying amounts in different preparations of GCX. Oligosaccharides were released from GCX by hydrazinolysis and separated by electrophoresis after reductive amination to 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS). Bands comigrating with standards containing 11, 12, 16, and 17 sugar residues were detected. Thus, the GCX is a complex structure composed of oligosaccharides, probably linked to a peptide.
覆盖血吸虫尾蚴的糖萼(GCX)是一种具有免疫调节特性的复杂分子。在此,我们使用鳗鲡凝集素纯化了毫克量的GCX,该凝集素可与覆盖尾蚴体和尾部的GCX结合。通常,用苯酚提取1000万个尾蚴,透析后在琼脂糖2B-CL柱上进行色谱分离。平均有39毫克总碳水化合物在空体积附近洗脱,其中31毫克类糖原物质通过凝集素亲和色谱进一步分离。通过成分分析、对淀粉酶消化的敏感性及其核磁共振光谱确定了其身份。凝集素结合的GCX用0.1 M岩藻糖洗脱,最终碳水化合物产量为5.3毫克。岩藻糖占总GCX碳水化合物的40%,还存在少量但大致等量的半乳糖、葡糖胺和半乳糖胺。核磁共振数据表明氨基糖是N-乙酰化的。葡萄糖也存在,但在不同的GCX制剂中含量不同。寡糖通过肼解从GCX中释放出来,并在还原胺化生成8-氨基萘-1,3,6-三磺酸(ANTS)后通过电泳分离。检测到与含有11、12、16和17个糖残基的标准品共迁移的条带。因此,GCX是一种由寡糖组成的复杂结构,可能与一种肽相连。