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Biochemistry. 1996 Nov 26;35(47):14947-57.

Mechanism of membrane permeabilization by sticholysin I, a cytolysin isolated from the venom of the sea anemone Stichodactyla helianthus.

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Centro di Fisica degli Stati Aggregati, Consiglio Nazionale delle Richerche-Istituto Trentino di Cultura, Povo (Trento), Italy.


Actinaria cytolysins are very potent basic toxins isolated from the venom of sea anemones, which are supposed to exert their toxic activity through formation of oligomeric pores in the host plasma membrane. To gain insight into their mechanism of action, the interaction of Stichodactyla helianthus sticholysin I (St-I) with lipid bilayers was studied. St-I increased the permeability of calcein-loaded lipid vesicles composed of different phospholipids. The rate of permeabilization improved when sphingomyelin (SM) was introduced into phosphatidylcholine (PC) vesicles, reaching an optimum value at equimolar concentrations of these two phospholipids. It was also a function of the pH, showing a local maximum of activity between pH 8 and 9 and a marked decrease at pH 10 and 11. Under optimal conditions (e.g., PC:SM 1:1, pH 8, toxin to vesicle ratio < 200), most of the toxin is bound to the lipid phase. The reduced toxin effect at low and high SM content, or at high pH, is principally due to a decreased toxin binding. From the dose dependence of the permeabilization, at constant lipid concentration, it was inferred that St-I increases membrane permeability by forming oligomeric pores comprising at least three cytolysin monomers. The involvement of oligomers was also suggested by the dependence of calcein release on the vesicle concentration at constant toxin dose. In fact, the time course of dye release was well described under all circumstances by a kinetic model which assumes that trimerization leads to a conductive pore. All the relevant equilibrium and rate constants were derived. Addition of St-I to one side of a planar lipid membrane increased the conductivity of the film in discrete steps of defined amplitude, indicating the formation of ion channels. The dose dependence of this effect was the same as with LUV. The channel was cation-selective and its conductance suggested a functional radius of about 1.0 nm, consistent with the size of the lesion previously observed in red blood cells. Pores exhibited rectification and voltage-dependent gating.

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