Cellular location of alpha-hemolysin in Staphylococcus aureus

Protoplast fractionation techniques were used to prepare subcellular fractions of three strains of Staphylococcus aureus for the analysis of alpha-hemolysin activity. More than 90% of the cellular hemolytic activity was localized in the periplasm, only a trace was detected in the cytoplasm, and none was found in either plasma membranes or mesosomes. This cellular portion constituted approximately 4 to 10% of the total hemolytic activity of the culture. The erythrocyte lysis spectrum and Sephadex G-100 filtration studies indicated that the cellular hemolysin closely resembles the extracellular form.