PURIFICATION AND SOME PROPERTIES OF AEROMONAS-HYDROPHILA HEMOLYSIN
PURIFICATION AND SOME PROPERTIES OF AEROMONAS-HYDROPHILA HEMOLYSIN
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DOI：
10.1128/iai.46.1.122-127.1984
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发表时间：
1984-01-01
影响因子：
3.1
通讯作者：
SAKAGUCHI, G
中科院分区：
文献类型：
--
作者：
ASAO, T;KINOSHITA, Y;SAKAGUCHI, G
一种由A.采用酸沉淀和季铵乙基葡聚糖凝胶层析法对从腹泻病人中分离的大肠杆菌进行纯化。溶血素的MW通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳[SDS-PAGE]估计为50，000，通过Sephadex G-100凝胶过滤估计为48，000。在pH 4.0的聚丙烯酰胺凝胶电泳和SDS-PAGE中，溶血素迁移为单一条带;在pH 9.4的电泳和薄层等电聚焦显示多条带。结果可能表明溶血素的电荷异构体。纯化的溶血素对兔红细胞具有134溶血单位/μ g蛋白质的溶血活性。它导致幼年小鼠肠道中液体积聚，并导致兔子结扎回肠袢。纯化的溶血素对Vero细胞也有细胞毒性，对小鼠也有致死毒性。所有这些生物活性在56 ℃下加热5分钟时丧失。C.显然，A。大肠杆菌溶血素是一种细胞毒性肠毒素。
A hemolysin produced by a strain of A. hydrophila isolated from a patient with diarrhea was purified by acid precipitation and quarternary aminoethyl-Sephadex chromatography. The MW of the hemolysin was estimated at 50,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis [SDS-PAGE] and at 48,000 by Sephadex G-100 gel filtration. In polyacrylamide gel electrophoresis at pH 4.0 and SDS-PAGE, the hemolysin migrated as a single band; electrophoresis at pH 9.4 and thin-layer isoelectric focusing demonstrated multiple bands. The results may indicate charge isomers of the hemolysin. The purified hemolysin had a hemolytic activity of 134 hemolytic units/.mu.g of protein on rabbit erythrocytes. It caused fluid accumulation in infant mouse intestines and rabbit ligated ileal loops. Purified hemolysin also elicited cytotoxicity to Vero cells and lethal toxicity to mice. All these biological activities were lost on heating for 5 min at 56.degree. C. Evidently, A. hydrophila hemolysin is a cytotoxic enterotoxin.