五步蛇毒毒中蛋白凝集素的发现及其凝集特性的研究

THE DISCOVERY OF ATOXIC PROTEIN AGGLUTININ FROM FIVE PACE SNAKE VENOM AND THE STUDY OF ITS AGGREGATION PROPERTIES

发现尖吻蝮蛇(Agkistrodon acutus)粗毒对动物细胞有凝集作用。顺序用DEAE—Sephadlex A-50,CM Sephadex C25柱层析,从尖吻蝮蛇毒中分离纯化出一种毒蛋白凝集素。经分子量,等电点和出血活性等实验测定确定该凝集素就是徐询等人早期分离、纯化的出血毒素Ⅲ(AaHⅢ)。这种凝集素对许多动物细胞都有凝集作用,例如脾淋巴细胞、HeLa细胞、人肾成纤维细胞和草鱼(Ctenoph argngodon idellus)。但对组细胞和血小板无作用。和植物凝集素PHA相比AaHⅢ产生细胞的凝集块体积要大些。对淋巴细胞起凝集作用的最低浓度为2.5ug/ml。能使HeLa细胞和淋巴细胞凝集在一起。如预先加肝素和溶菌酶可以抑制对淋巴细胞的凝集怍用。用溶菌酶(2mg/ml)和AaHⅢ(1mg/ml)在37℃′下保温2小时AaHⅢ的出血活性仍然存在。

It was discovered that the crude Venom of Agkistrodon acutus can aggregate animal cells. We purified a toxic protein aggiutinln from A. aeutus venom sequencely by DEAE—Sephadex A—50 and CM-Sephadex C—25. This agglutinin is jest the hemorrhaginⅢ previously purified by Xu et al. Its properties, such as molecular weight, isoelectrie poin and hemorrhagic activity, were the same as hemorrhaginⅢ (AaHⅢ)、It cant aggregate many animal cells, including lymphocytes, Hela cells, fibroblast cells derived from human kidney and Cells derived from the kidney of Grass Carp (ctenopharyngodon idellus). But it had no effects to erythrocytes and platetes. As compared with PHA, the cell clots formed by the aggregation action of AaHⅢ were larger. The lowest concentration at which AaHⅢ can also induce lymphocytes aggregate was 2.5ug/ml. It can make Hela cells and lymphocytes aggregate together. The aggregation action may be inhibited by heparin and lysozyme which were added before AaHⅢ. Its hemorrhagic activity remained after AaHⅢ (Img/ml) was incubated with lysozyme(2mg/ml) for 2hr at 37℃.