Multiple forms of staphylococcal alpha-toxin.
A group of proteins was readily extracted at neutrality from trichloroacetic acid precipitates of staphylococcal culture filtrate supernatants, while alpha-toxin was dissolved and activated by treating the precipitate with 8 M urea, with acidic buffers or by heating to 90-100 degrees C at neutrality. Heat activation of the precipitate produced a relatively pure alpha-toxin with a molecular weight of 39,000. alpha-Toxin was eluted together with three other proteins on hydroxyl apatite chromatography, and evidence was obtained for an association between the four proteins. On isoelectric focusing a haemolytic fraction was obtained at pH 6.2, probably due to acid activation of the precipitate formed at the cathodic end of the column. The alpha-haemolytic fractions with pI's of 7.4 and 8.6 were shown to consist of alpha-toxin only when analyzed by acrylamide electrophoresis in the presence of sodium dodecyl sulphate. The haemolytic component with a pI of 9.2 contained two additional components of molecular weights of 27,500 and 18,000. Chromatography of this material on Sephadex G-200 showed that alpha-toxin and the two proteins appeared as a high molecular complex.
D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006461:Hemolysis; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D008970:Molecular Weight; D011506:Proteins / Q000032:analysis; D012967:Sodium Dodecyl Sulfate; D013211:Staphylococcus aureus; D014118:Toxins, Biological / Q000302:isolation & purification*; D014508:Urea
D011506:Proteins; D014118:Toxins, Biological; D012967:Sodium Dodecyl Sulfate; D014508:Urea