Studies With Staphylococcal Toxins: Iv. The Purification and Metallic Requirements of Specific Hemolysins

A method is described whereby specific staphylococcal hemolysins may be concentrated to provide a 600-fold increase in potency as compared with the initial culture filtrate.The progressive purification of the hemolysins was conveniently followed by the determination of the quantity of hemin (in μg.) freed from a standard suspension of erythrocytes per μg. of protein in the toxic preparation. All preparations of the electrophoretically pure hemolysins were protein in nature, but in order to effect hemolysis, preparations of similar electrophoretic mobility obtained from representatives of two different phage groups (IV and II) showed, respectively, an absolute requirement and no requirement for the presence of specific divalent cations. The initial culture filtrate was dermonecrotic and lethal to test animals. The "purified" hemolysins had neither of these properties.Two "purified" hemolysins that showed differences in biological, electrophoretic, and chemical properties appeared to be serologically homologous. The properties of the "purified" hemolysins from either strain differed from the more generally accepted properties of alpha- and beta-hemolysins even though the culture filtrate seemed to contain these two classically recognized lysins.