中国鲎凝集素的分离纯化及性质研究

 经N-乙酰氨基葡萄糖交联琼脂糖亲和层析及以交联琼脂糖介质的高效液相分子筛层析,从中国鲎细胞溶解物中分离纯化了一种凝集素,其活性比原料鲎试剂提高128倍。鲎凝集素SDS电泳时表现出分子量为69000,和72000的二个亚基。N-乙酰氨基葡萄糖、D-半乳糖,D-甘露糖及岩藻糖等对鲎凝集素凝集鸡红细胞的活性有显著抑制作用,加热60℃,10分钟可使凝集素活性基本丧失。CaCl_2为凝集素活性所必需。鲎凝集素与肺炎球菌C多糖有沉淀反应。

Isolation, Purification and Characterization of a Lectin From the Chinese Horseshoe Crab, Tachypleus Tridendatus

A lectin was isolated from the hemocyte of the Chinese horseshoe crab, Tachypleus tridendatus, by affinity chromatography on a N-acetyl-D-glucosamine agarose column. The sample isolated from affinity chromatography was further applied to a HPLC column packed with cross-linked agarose beads as molecular sieve for fractionation into three components. The major component had a 128 fold increase in agglutinating activity as compared with the starting material. On SDS-PAGE, the lectin appeared to be composed of two heterogeneous subunits with molecular weight of 69000 and 72000 daltons respectively. Its activity was specifically inhibited by N-acetyl-D-glucosamine and D-galactose. Heating at 60℃ for 10 min inactivated the lectin. The lectin can be activated by calcium chloride and forms preciptate with the C-polysaccharide of pneumococcus.