Sugahara T, Takahashi T, Yamaya S, Ohsaka A
Jpn J Med Sci Biol. 1976 Oct;29(5):255-63.
Highly purified alpha-toxin (phospholipase C) of Clostridium perfringens prepared by affinity chromatography on agarose-linked egg-yolk lipoprotein induced the in vitro aggregation of platelets of an irreversible type. The aggregation started after a time lag, the length of which depended on the concentration of the toxin; the reciprocal of the time lag was found to be directly proportional to the toxin concentration. Using this assay method, we demonstrated that the platelet-aggregating activity of alpha-toxin reached minimum at around 70 C but heating at higher temperatures inactivated it to a lesser extent; the same anomaly in heat inactivation was observed with phospholipase C activity possessed by the toxin. By subjecting purified alpha-toxin to isoelectric focusing, four molecular forms were isolated, all of which were associated with both the platelet-aggregating and phospholipase C activities. From all these results we concluded that the entity responsible for the platelet-aggregating activity is identical with alpha-toxin (phospholipase C).
通过琼脂糖连接的蛋黄脂蛋白亲和层析制备的产气荚膜梭菌高度纯化的α毒素（磷脂酶C）可在体外诱导血小板发生不可逆类型的聚集。聚集在一段延迟时间后开始，延迟时间的长短取决于毒素的浓度；发现延迟时间的倒数与毒素浓度成正比。使用这种检测方法，我们证明α毒素的血小板聚集活性在70℃左右达到最小值，但在较高温度下加热使其失活的程度较小；毒素所具有的磷脂酶C活性在热失活方面也观察到同样的异常。通过对纯化的α毒素进行等电聚焦，分离出四种分子形式，所有这些形式都与血小板聚集活性和磷脂酶C活性相关。从所有这些结果我们得出结论，负责血小板聚集活性的实体与α毒素（磷脂酶C）相同。
