Suzuki K, Une T, Yamazaki M, Takeda T
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Toxicon. 1990;28(9):1019-28. doi: 10.1016/0041-0101(90)90140-3.
A hemolytic protein which causes diarrhea and death to mice was purified from the fruit bodies of a poisonous mushroom species Rhodophyllus rhodopolius (Fries) by DEAE-cellulose column chromatography, ammonium sulfate precipitation, gel filtration on Sephadex G-200, and DEAE-Sephadex A-25 column chromatography. The mol. wt of the purified hemolysin was estimated to be 40,000 by SDS-polyacrylamide slab gel electrophoresis. The hemolytic activity of the purified hemolysin was destroyed by heating at 60 degrees C for 10 min, and partially reduced by pepsin, papain and 2-mercaptoethanol. Cholesterol and phosphatidylcholine did not inhibit the activity. The hemolysin was unstable below pH 7.0 but stable at pH 8.0. The optimal pH for hemolysis was 6.0. Hemolysis did not occur below 4 degrees C even though the hemolysin bound to the erythrocyte. Mouse, chicken, rat, horse and human erythrocytes were sensitive in this order, but sheep and cow erythrocytes were not lysed by the toxin.
通过DEAE - 纤维素柱层析、硫酸铵沉淀、Sephadex G - 200凝胶过滤和DEAE - Sephadex A - 25柱层析，从有毒蘑菇红褶孔菌（Rhodophyllus rhodopolius (Fries)）的子实体中纯化出一种能导致小鼠腹泻和死亡的溶血蛋白。经SDS - 聚丙烯酰胺平板凝胶电泳估计，纯化的溶血素分子量为40,000。纯化的溶血素在60℃加热10分钟后溶血活性被破坏，经胃蛋白酶、木瓜蛋白酶和2 - 巯基乙醇处理后活性部分降低。胆固醇和磷脂酰胆碱不抑制其活性。溶血素在pH 7.0以下不稳定，但在pH 8.0时稳定。溶血的最佳pH为6.0。即使溶血素与红细胞结合，在4℃以下也不发生溶血。小鼠、鸡、大鼠、马和人的红细胞按此顺序敏感，但绵羊和牛的红细胞不被该毒素裂解。
