红芸豆红细胞凝集素单体的分离纯化和性质研究  

Purification and Partial Characterization of Erythrohemagglutinin from Phaseolus Vulgaris

作　　者：何涛[1] 张涛[2] 吴昌英[1] 王亮[1] 单晓雪[1] 罗琴[1] 

机构地区：[1]成都医学院基础医学院 [2]成都医学院基础医学院基础医学院生物技术实验室,成都610083

出　　处：《成都医学院学报》2008年第2期107-110,119,共5页Journal of Chengdu Medical College

基　　金：成都医学院实验室开放基金资助项目(项目编号)JC200816

摘　　要：目的研究一种从红芸豆中提取红细胞凝集素单体(PHA-E)的新方法。方法红芸豆经过浸提,离子交换树脂分离,分子筛层析纯化后得到PHA-E样品。采用电泳法测定其纯度、分子量和等电点。用2%人细胞悬液测定样品凝集红细胞的能力和影响凝血的因素,使用硫酸苯酚法测定其糖含量。结果经PAGE分析PHA-E样品为单带电泳纯,SDS-PAGE上显示亚基分子量为32kD,等电点为6.5.样品使人红细胞50%凝集的蛋白质最低浓度为4μg/ml左右。单糖不影响PHA-E凝血活力,EDTA抑制其凝血活力,Zn2+促进其凝血。糖含量为8.1%.Objective To study a new method for Erythrohemagglutinin （PHA- E） production from Phaseolus vulgaris. Methods PHA - E was purified from the excellent seeds of Phaseolus vulgaris by extraction, ion exchange chromatography and final gel filtra- tion chromatography. The purity, the molecular weight and the isoelectric point of purified PHA - E were determined by electrophoresis. The ability and the influencing factor of agglutination were determined by 2 % erythrocytes of human. Conclusion The purified PHA - E was single PAGE outhne and had apparent subunit molecular weights of 32 kD by SDS - polyacrylamide gel electrophoresis. Isoelectric point of the PHA - E was 6.5. PHA - E can promote agglutination of human erythrocytes at 4 μg/ml, but its activity was not inhibited by any monosaccharide. EDTA can inhibit the abihty of agglutination, and Zn^2 ＋ can accelerate it. The sugar content of PHA - E was 8.1%.

关 键 词：红细胞凝集素 分离纯化 凝血 性质研究 

分 类 号：Q513.2[生物学—生物化学]