Purification and characterization of Clostridium perfringens delta-toxin.
Delta-toxin, an extracellular hemolysin released by Clostridium perfringens type C, was purified from culture supernatant fluid by sequential ammonium sulfate precipitation, thiol-Sepharose gel chromatography, isoelectric focusing, and Sephadex G-75 gel filtration. The purified preparation had a specific activity of 320,000 hemolytic units per mg of protein and was homogeneous, as determined by immunochemical and electrophoretic tests. This toxin was characterized as a single polypeptide chain composed of 391 amino acid residues, 30% of which were hydrophobic. The molecular weight was found to be 42,000, and the isoelectric point was pH 9.1. Delta-toxin appeared to be amphiphilic by charge shift electrophoresis in a three-detergent system. It was immunogenic in rabbits and lethal to mice at a dose of 0.12 micrograms. The lytic activity of delta-toxin was restricted to erythrocytes of even-toed ungulates (sheep, goats, and pigs). This activity was inhibited by GM2 ganglioside but not by other gangliosides, cholesterol, lecithin, or sphingomyelin.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001427:Bacterial Toxins / Q000302:isolation & purification*; D003016:Clostridium perfringens / Q000276:immunology*; D006460:Hemolysin Proteins / Q000302:isolation & purification* / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D007526:Isoelectric Point; D008055:Lipids / Q000494:pharmacology; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D012995:Solubility
D000596:Amino Acids; D001427:Bacterial Toxins; C029352:Clostridium perfringens delta-toxin; D006460:Hemolysin Proteins; D008055:Lipids; D008670:Metals