On the mechanism of membrane damage by Staphylococcus aureus alpha-toxin.
Rabbit or human erythrocytes lysed with Staphylococcus aureus alpha-toxin were solubilized with Triton X-100, and the toxin was subsequently isolated by gel chromatography, sucrose density gradient centrifugation, and reincorporation into liposomes. In the presence of Triton X-100, the toxin exhibited a sedimentation coefficient of 11S and eluted at a position between those of IgG and alpha 2-macroglobulin in gel chromatography. A single polypeptide subunit of 34,000 mol wt was found in SDS PAGE. In the electron microscope, ring-shaped or cylindrical structures were observed, 8.5-10 nm in diameter, harboring central pits or channels 2-3 nm in diameter. An amphiphilic nature of these structures was evident from their capacity to bind lipid and detergent, aggregation in the absence of detergents, and low elutability from biological and artificial membranes through ionic manipulations. In contrast to the membrane-derived form of alpha-toxin, native toxin was a water-soluble, 34,000 mol wt, 3S molecule, devoid of an annular structure. Because studies on the release of radioactive markers from resealed erythrocyte ghosts indicated the presence of circumscribed lesions of approximately 3-nm effective diameter in toxin-treated membranes, the possibility is raised that native alpha-toxin oligomerizes on and in the membrane to form an amphiphilic annular complex that, through its partial embedment within the lipid bilayer, generates a discrete transmembrane channel.
D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects*; D006460:Hemolysin Proteins; D006801:Humans; D007473:Ion Channels; D008051:Lipid Bilayers; D008081:Liposomes; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D008970:Molecular Weight; D009498:Neurotoxins / Q000494:pharmacology*; D012756:Sheep
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; D008081:Liposomes; D046911:Macromolecular Substances; D008565:Membrane Proteins; D009498:Neurotoxins; C021091:staphylococcal alpha-toxin