Lind I, Ahnert-Hilger G, Fuchs G, Gratzl M
Abteilung Anatomie und Zellbiologie, Universität Ulm, Federal Republic of Germany.
Anal Biochem. 1987 Jul;164(1):84-9. doi: 10.1016/0003-2697(87)90371-x.
Crude alpha-toxin was produced by Staphylococcus aureus, strain Wood 46. The amount of exotoxin was monitored during growth and all subsequent purification steps by determination of its hemolytic activity against rabbit erythrocytes. The culture supernatant was treated with ammonium sulfate (75% saturation). The resulting precipitate was dialyzed and subjected to cation-exchange chromatography. The fractions containing the hemolytic activity were further purified by gel chromatography. The final product was enriched by a factor of 8.5 compared to the crude toxin. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified toxin exhibited one major band. It caused the release of 86Rb+ and ATP from rat insulinoma (RIN A2) as well as pheochromocytoma cells (PC12) in culture, indicating efficient permeabilization of their plasma membranes for small molecules.
金黄色葡萄球菌伍德46株产生了粗制α毒素。在生长过程以及随后所有的纯化步骤中，通过测定其对兔红细胞的溶血活性来监测外毒素的量。培养上清液用硫酸铵（75%饱和度）处理。所得沉淀物进行透析并进行阳离子交换色谱分析。含有溶血活性的级分通过凝胶色谱进一步纯化。最终产物比粗毒素浓缩了8.5倍。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中，纯化后的毒素呈现出一条主要条带。它导致培养中的大鼠胰岛素瘤（RIN A2）细胞以及嗜铬细胞瘤细胞（PC12）释放86Rb+和ATP，表明其质膜对小分子有高效的通透作用。
