Oligomer formation of staphylococcal alpha-toxin analyzed by electron microscopy and image processing

Abstract

The 12S oligomeric form of Staphylococcus aureus alpha-toxin has been studied with electron microscopy after incubation of the toxin with membrane preparations or liposomes. The target material originated from human platelets. Different electron microscopic preparation techniques were used including negative staining, freeze-fracture and vitrification in liquid ethane. Analysis of micrographs with image processing methods revealed two groups of ring-like structures corresponding to alpha-toxin oligomers. One form measured 75 A in diameter and had a high stain density in the central protein deficient part while the other was larger with a diameter of 100 A and less stain accumulation in the center. The conditions under which the latter were formed suggest that this corresponds to an inactive loosely-bound form of the toxin. The high stain density in the smaller particle is consistent with the presence of a penetrating pore in this structure.