Mechanism of Magainin 2a Induced Permeabilization of Phospholipid Vesicles

Authors

Earl Grant, Brooke Army Medical Center
Troy J. Beeler, Uniformed Services University of the Health Sciences
Kenneth M. P. Taylor, Uniformed Services University of the Health Sciences
Kenneth Gable, Uniformed Services University of the Health Sciences
Mark A. Roseman, Uniformed Services University of the Health Sciences

Date of this Version

1992

Comments

Published in Biochemistry (1992) 31, 9912-9918

Abstract

The magainins, peptide antibiotics secreted by the frog Xenopus laevis, have previously been shown to permeabilize phospholipid vesicles. To elucidate the mechanism of permeabilization, we have conducted detailed kinetic studies of magainin 2 amide (mgn2a)hduced release of 6-carboxyfluorescein from vesicles of phosphatidylserine. The results show that dye release occurs in (at least) two stages-an initial rapid phase, with t_1/2 ≈ 3 s, followed by a much slower phase that approaches zero leakage rate before all the dye is released. Light-scattering studies showed that mgn2a does not cause gross changes in vesicle structure. The peptide was found to rapidly equilibrate between vesicles; this was demonstrated by determining a binding isotherm for the peptidelipid interaction, and by showing that addition of unloaded vesicles rapidly quenches peptide-induced leakage from loaded vesicles. Transient dye release in the presence of an equilibrating peptide can be explained in two ways: (1) the peptide exists only transiently in an active form; (2) the vesicles are only transiently leaky. Preincubation of mgn2a at assay concentrations in buffer alone or with unloaded vesicles did not inactivate the peptide. Therefore, rapid leakage is probably due to transient destabilization of the vesicle upon addition of mgn2a.