Manohar M, Maheswaran S K, Frommes S P, Lindorfer R K
J Bacteriol. 1967 Jul;94(1):224-31. doi: 10.1128/jb.94.1.224-231.1967.
Crude and purified staphylococcal alpha-toxin were used to demonstrate that the platelet-damaging effect of crude alpha-toxin represents a fifth activity of the alpha-toxin molecule. The homogeneity of the purified toxin employed was demonstrated by ultracentrifugation, Ouchterlony, and immunoelectrophoretic methods. Continuous-flow electrophoretic migration studies demonstrated under a variety of conditions that the platelet-damaging and the alpha-hemolytic activities migrated as a unit. Fractionation studies with the use of Sephadex G-100, carboxymethyl cellulose, and diethylaminoethyl cellulose failed to separate these two activities. Further, when alpha-toxin of demonstrated purity and crude toxin were adjusted to the same hemolytic activity, they possessed the same platelet-damaging activity. In addition, heat-reactivation studies with crude alpha-toxin revealed that the platelet-damaging effect was inactivated and reactivated in parallel with alpha-hemolytic activity. Comparable studies with purified alpha-toxin showed parallel inactivation of both activities at 60 C. Additional heating at 100 C failed to reactivate either activity. Electron micrographs revealed that purified alpha-toxin produced distinct degenerative changes in rabbit platelets. These studies also provided definite evidence that purified alpha-toxin has a damaging effect on human platelets. Monovalent alpha-antisera prevented platelet damage.
使用粗制和纯化的葡萄球菌α毒素来证明粗制α毒素对血小板的损伤作用代表了α毒素分子的第五种活性。通过超速离心、双向免疫扩散和免疫电泳方法证明了所用纯化毒素的均一性。连续流动电泳迁移研究表明，在各种条件下，血小板损伤活性和α溶血活性作为一个整体迁移。使用葡聚糖G-100、羧甲基纤维素和二乙氨基乙基纤维素进行的分级分离研究未能分离这两种活性。此外，当将已证明纯度的α毒素和粗毒素调整到相同的溶血活性时，它们具有相同的血小板损伤活性。此外，对粗制α毒素进行的热再激活研究表明，血小板损伤作用与α溶血活性同时失活和再激活。对纯化α毒素进行的类似研究表明，在60℃时两种活性同时失活。在100℃进一步加热未能使任何一种活性再激活。电子显微镜照片显示，纯化的α毒素在兔血小板中产生明显的退行性变化。这些研究还提供了确凿的证据，证明纯化的α毒素对人血小板有损伤作用。单价α抗血清可防止血小板损伤。
