PURIFICATION AND CHARACTERIZATION OF A STAPHYLOCOCCAL EPIDERMOLYTIC TOXIN
PURIFICATION AND CHARACTERIZATION OF A STAPHYLOCOCCAL EPIDERMOLYTIC TOXIN
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DOI：
10.1128/iai.13.2.627-633.1976
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发表时间：
1976-01-01
影响因子：
3.1
通讯作者：
WUEPPER, KD
中科院分区：
文献类型：
--
作者：
DIMOND, RL;WUEPPER, KD
一种葡萄球菌外毒素，当注射到新生小鼠和人的皮肤中时会引起表皮炎，这种外毒素是通过常规的生物化学技术高度纯化的。对于金黄色葡萄球菌EV，表皮毒素是上清液培养液的主要蛋白组分。纯化的初始步骤是在pH9.0下在Pevikon中进行的区带电泳，pH9.0是α-葡聚糖的等电点。溶血性毒素，其保持在起源附近。含有表皮毒素但不含α-葡萄糖的级分毒素，然后在羧甲基-Sephadex C-50上进行阳离子交换层析以除去痕量污染物。一种主要的高度纯化的表皮坏死毒素在聚丙烯酰胺凝胶电泳中迁移为单一条带，在分析超滤中沉淀为单一组分，并在注射到家兔体内后引起单一沉淀抗体。还鉴定了较少量的第二种表皮炎毒素，其分子量和抗原性相同，但电泳行为与主要分子种类不同。表皮生长因子的分子量为28，600 ±。通过十二烷基硫酸钠-丙烯酰胺电泳测定为400 ± 0.40，和32，500 ± 0.40。120接近沉降平衡。
A staphylococcal exotoxin that causes epidermolysis when injected into the skin of the newborn mouse and man was highly purified by conventional biochemical techniques. With Staphylococcus aureus EV, the epidermolytic toxin was a major protein component of supernatant culture fluids. The initial step in purification was zone electrophoresis in Pevikon carried out at pH 9.0, the isoelectric point of .alpha.-hemolytic toxin which remained near the origin. Fractions containing the epidermolytic toxin, but free of .alpha.-toxin, were then subjected to cation exchange chromatography on carboxymethyl-Sephadex C-50 to remove trace contaminants. A major highly purified epidermolytic toxin migrated as a single band in polyacrylamide gel electrophoresis, sedimented as a single component in the analytical ultracentrifuge and elicited a single precipitating antibody after injection into rabbits. A smaller amount of a 2nd epidermolytic toxin, identical in molecular weight and antigenicity but differing in electrophoretic behavior from the major molecular species, was also identified. The epidermolytic factor had a molecular weight of 28,600 .+-. 400 by sodium dodecyl sulfate-acrylamide electrophoresis, and 32,500 .+-. 120 by approach to sedimentation equilibrium.