摘要
江苏菜豆经酸水(PH2.0)抽提,硫酸铵分级沉淀,分离植物血球凝集素(PHA-P),分子量为128000的糖蛋白,活性回收率在80％以上,PHA-P经SP-sephadexc-50离子交换层析,分成L_4,L_3E_1,L_2E_2,L_1E_3,和E_4同工凝集素。 L_4和E_4等电点为5.4和6.5。亚基分子量分别是31000和33000,并有类似的氨基酸组成。PAGE分析为单一蛋白带。红细胞凝集活性随电泳迁移速度的加快而增强,促淋性细胞分裂活性则减弱。E_4血凝活性受CalNAc,EDTA抑制和Zn^(++)的促进。
Phytohemagglutinin (PHA-P) from Jiangsu red kidney bean (Phaseolus vulgaris) was extracted in distilled water (pH2.0) and precipitated with ammonium sulfate.PHA-P is a glycoprotein with molecular weights is 128 000.Over 80% of the hemagglutinating activity was recovered.PHA-P resolves into L4, L3E1, L2E2, L1E3 and E4 isolectins by SP-Sephadex C-50 ion exchange chromatography, and all of them have apparent subunit molecular weights of 31000 and 33000 by SDS-polyacry-lamide gel electrophoresis.They also have similar amino acid compsition. Isoelectric point of the L4 and E4 were 5.4 and 6.5, respectively. The five separated isolectins migrate as single bands of increasing mobility in the electrophoresis. Along with the increase of migrating rate,erythroagglutinating activity has been strengthened,and the mitogenic activity has been weakened, Hemagglutination activity of E4 was inhibited by GalNAc, and EDTA but increased by Zn++.
关键词
菜豆
同工凝集素
分离
纯化
Phytohemagglutinin
Isolectin
Ion exchange chromatography
Purification
