Zitzer A O, Nakisbekov N O, Li A V, Semiotrochev V L, Muratkhodjaev J N, Krasilnikov O V
Middle Asian Antiplague Research Institute, Alma-Ata, Kazakhstan.
Zentralbl Bakteriol. 1993 Nov;279(4):494-504. doi: 10.1016/s0934-8840(11)80421-4.
A thermolabile extracellular entero-cytolysin (EC) from Vibrio cholerae non-O1 was purified by ammonium sulphate fractionation, DE-52 cellulose ion exchange chromatography, gel-filtration on Ultrogel AcA-44 and high performance liquid chromatography on a Mono Q. The purified EC had a molecular weight of 63 kD and an isoelectric point of 6.2. It was not inactivated by cholesterol or 5,5'-dithio-bis(2-nitrobenzoic acid), nor activated by dithiothreitol. EC had no immunological cross-reactivity with cholera toxin. The EC caused fluid accumulation in the intestines of infant rabbits, death of mice by intravenous injection, and increased vascular permeability in the paw oedema test in mice. V. cholerae non-O1 EC lysed erythrocytes from various species and cultured cells (CHO, L-929, L-41, HEp-2, Vero, MDCK and BHK-21). In contrast to cholera toxin, EC caused crude destruction of target cells. The EC caused hemolysis by a colloid-osmotic mechanism due to the formation of hydrophilic pores of 1.8-2.0 nm diameter in the cell membrane. This EC also was able to open pores in lipid membranes. The induced channels were anion-selective and had a diameter of 1.8-2.0 nm.
通过硫酸铵分级沉淀、DE-52纤维素离子交换色谱、Ultrogel AcA-44凝胶过滤和Mono Q高效液相色谱法，从非O1群霍乱弧菌中纯化出一种热不稳定的细胞外肠细胞毒素（EC）。纯化后的EC分子量为63 kD，等电点为6.2。它不会被胆固醇或5,5'-二硫代双（2-硝基苯甲酸）灭活，也不会被二硫苏糖醇激活。EC与霍乱毒素没有免疫交叉反应。EC可导致幼兔肠道积液、通过静脉注射使小鼠死亡，并在小鼠爪肿胀试验中增加血管通透性。非O1群霍乱弧菌EC可裂解来自各种物种的红细胞以及培养细胞（CHO、L-929、L-41、HEp-2、Vero、MDCK和BHK-21）。与霍乱毒素不同，EC会导致靶细胞的粗破坏。EC通过胶体渗透机制引起溶血，这是由于在细胞膜上形成了直径为1.8 - 2.0 nm的亲水性孔。这种EC还能够在脂质膜上形成孔。诱导形成的通道具有阴离子选择性，直径为1.8 - 2.0 nm。
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