CAJ | 학술논문

建立运用兔红细胞膜制备亲和树脂来纯化红芸豆中红细胞凝集素的方法.红芸豆经过浸提,(NH_4)_2SO_4沉淀,红细胞膜亲和树脂吸附、洗脱得到红细胞凝集素(PHA-E)试样.采用电泳法测定其纯度、相对分子质量和等电点.用体积分数2%的兔红细胞悬液测定试样凝血活力及影响凝血因素.经PAGE分析PHA-E试样为单带,SDS-PAGE分析显示亚基相对分子质量为3.2×104,等电点为6.5.研究发现,促使50%兔红细胞产生凝集的试样蛋白质最低质量浓度为4 μg/mL,单糖不影响PHA-E凝血活力,EDTA抑制其凝血活力,Zn~(2+)促进其凝血.
건립운용토홍세포막제비친화수지래순화홍예두중홍세포응집소적방법.홍예두경과침제,(NH_4)_2SO_4침정,홍세포막친화수지흡부、세탈득도홍세포응집소(PHA-E)시양.채용전영법측정기순도、상대분자질량화등전점.용체적분수2%적토홍세포현액측정시양응혈활력급영향응혈인소.경PAGE분석PHA-E시양위단대,SDS-PAGE분석현시아기상대분자질량위3.2×104,등전점위6.5.연구발현,촉사50%토홍세포산생응집적시양단백질최저질량농도위4 μg/mL,단당불영향PHA-E응혈활력,EDTA억제기응혈활력,Zn~(2+)촉진기응혈.
A method for erythrohemagglutinin(PHA-E)production from Phaseolus vulgaris and its purification by rabbit erythrocyte membrane affinity resin was established.PHA-E was purified from the seeds of Phaseolus vulgaris by extraction,(NH_4)_2SO_4 precipitation,adsorption by erythrocyte membrane affinity resin, and elution.The molecular weight and the isoelectric point of purified PHA-E were determined. The ability and the influencing factor of the agglutination were determined by 2% erythrocytes of human. The purified PHA-E showed single PAGE band and the apparent molecular weights of the subunit was 3.2×104 by SDS-PAGE and the isoelectric point of the PHA-E was 6.5.PHA-E could promote agglutination of human erythrocytes at 4 μg/mL,and its activity was not inhibited by monosaccharide.EDTA inhibited the agglutination ability,and Zn~(2+) accelerated its blood coagnlation.