Pore-formation by Escherichia coli hemolysin (HlyA) and other members of the RTX toxins family.

Escherichia coli hemolysin (HlyA) is a major cause of E. coli virulence. It lyses erythrocytes by a colloid osmotic shock due to the formation of hydrophilic pores in the cell wall. The size of these channels can be estimated using osmotic protectant of increasing dimensions. To show that the formation of pores does not depend critically on the osmotic swelling we prepared resealed human erythrocyte ghosts loaded with a fluorescent marker. When attacked by HlyA the internal marker was released, indicating the formation of toxin channels so large as to let it through. The channels can be directly demonstrated also in purely lipidic model systems such as planar membranes and unilamellar vesicles, which lack any putative protein receptor. HlyA has been recognised as a member of a large family of exotoxins elaborated by Gram-negative organisms including Proteus, Bordetella, Morganella, Pasteurella and Actinobacillus. These toxins have quite different target cell specificity and in many cases are leukocidal. When tried on planar membranes however, even specific leukotoxins open channels not dissimilar from those formed by HlyA, suggesting this might be a common step in their action. Comparison of the hydrophobic properties of six members of the toxin family indicates the presence of a conserved cluster of ten contiguous amphipathic helixes, located in the N-terminal half of the molecule, which might be involved in channel formation.