Staphylococcal beta-hemolysin. II. Phospholipase C activity of purified beta-hemolysin.
Sheep erythrocyte ghosts released water-soluble organic phosphorus when treated with purified beta-hemolysin. Phospholipid analysis demonstrated that sphingomyelin accounted for 53% of the phospholipids present in sheep erythrocytes. Purified beta-hemolysin showed phospholipase C activity when purified ox brain or sheep erythrocyte sphingomyelin was used as substrate. Such studies have also revealed that the disappearance of sphingomyelin from the reaction mixture was accompanied by a comparable increase in the concentration of phosphoryl choline. Thin-layer chromatography of phospholipids, extracted from sheep erythrocytes which had been exposed to beta-hemolysin, demonstrated that sphingomyelin was rapidly degraded. Activators of beta-hemolysin, such as Mg(++), enhanced the release of organic phosphorus from erythrocyte ghosts and from sphingomyelin. Inhibitors of beta-hemolysin, such as ethylenediaminetetraacetic acid, p-chloromercuribenzoate, and iodoacetamide, also inhibited the release of organic phosphorus from erythrocyte ghosts and from sphingomyelin. These studies strongly suggested that beta-hemolysin enzymatically degraded the sphingomyelin of the erythrocyte membrane. Such degradation probably resulted in the eventual lysis of the erythrocyte.
D000818:Animals; D001923:Brain Chemistry; D002729:Chloromercuribenzoates / Q000494:pharmacology; D002794:Choline / Q000032:analysis; D002855:Chromatography, Thin Layer; D004492:Edetic Acid / Q000494:pharmacology; D004912:Erythrocytes / Q000032:analysis / Q000378:metabolism; D006460:Hemolysin Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D066298:In Vitro Techniques; D007461:Iodoacetates / Q000494:pharmacology; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000378:metabolism; D010713:Phosphatidylcholines / Q000032:analysis; D010714:Phosphatidylethanolamines / Q000032:analysis; D010740:Phospholipases / Q000378:metabolism*; D010743:Phospholipids / Q000032:analysis; D010758:Phosphorus / Q000032:analysis; D012756:Sheep; D013109:Sphingomyelins / Q000032:analysis / Q000378:metabolism; D013210:Staphylococcus / Q000201:enzymology*; D013696:Temperature
D002729:Chloromercuribenzoates; D006460:Hemolysin Proteins; D007461:Iodoacetates; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D013109:Sphingomyelins; D010758:Phosphorus; D004492:Edetic Acid; D010740:Phospholipases; D010447:Peptide Hydrolases; D008274:Magnesium; D002794:Choline