Molecular dynamics of the cyclic lipodepsipeptides' action on model membranes: effects of syringopeptin22A, syringomycin E, and syringotoxin studied by EPR technique.

Interaction of pore-forming toxins, syringopeptin22A (SP22A), syringomycin E (SRE) and syringotoxin (ST), with model membranes were investigated. Liposomes were prepared from saturated phospholipids (DPPC or DMPC) or from binary mixtures of DPPC with varying amount of DOPC or cholesterol. The effects of the three toxins on the molecular order and dynamics of the lipids were studied using electron paramagnetic resonance (EPR) techniques. SP22A was the most-, SRE less-, and ST the least effective to increase the ordering and to decrease the rotational correlation time of the lipid molecules. The effects were more pronounced: (a) on small unilamellar vesicles (SUVs) than on multilamellar vesicles (MUVs); (b) on pure DPPC than on DPPC-cholesterol or DPPC-DOPC mixtures. Fluidity changes, determined from EPR spectra at different concentrations of the toxin, suggested the shell structure of the lipid molecules in pore formation. EPR spectra observed at different depth of the hydrocarbon chain of the lipid molecules implied an active role of the lipid molecules in the architecture of the pores created in the presence of the three toxins. Temperature dependence of the fluidity of the SUVs treated with toxins has shown an abrupt and irreversible change in the molecular dynamics of the lipid molecules at a temperature close to the pretransition, depending on the toxin species and the lipid composition. Coalescence and aggregation of the SUVs were proposed as the origin of this irreversible change.

Syringotoxin pore formation and inactivation in human red blood cell and model bilayer lipid membranes.

The effect of syringotoxin (ST), a member of the cyclic lipodepsipeptides family (CLPs) produced by Pseudomonas syringae pv. syringae on the membrane permeability of human red blood cells (RBCs) and model bilayer lipid membranes (BLMs) was studied and compared to that of two recently investigated CLPs, syringomycin E (SRE) and syringopeptin 22A (SP22A) [Biochim. Biophys. Acta 1466 (2000) 79 and Bioelectrochemistry 52 (2000) 161]. The permeability-increasing effect of ST on RBCs was the least among the three CLPs. A time-dependent ST pore inactivation was observed on RBCs at 20 and 37 degrees C but not at 8 degrees C. From the kinetic model worked out parameters as permeability coefficient of RBC membrane for 86Rb(+) and pores mean lifetime were calculated. A shorter pores mean lifetime was calculated at 37 degrees C then at 20 degrees C, which gave us an explanation for the unusual slower rate of tracer efflux measured at 37 degrees C then that at 20 degrees C. The results obtained on BLM showed that the pore inactivation was due to a decrease in the number of pores but not to a change of their dwell time or conductance.