Peptidyl-glycine-leucine-carboxyamide (PGLa), isolated from granular skin glands of Xenopus laevis, is practically devoid of secondary structure in aqueous solution and in the presence of zwitterionic phospholipids, when added exogenously, but adopts an alpha-helix in the presence of anionic lipids. The peptide was shown to exhibit antifungal activity and to have antimicrobial activity towards both Gram-negative and Gram-positive bacteria. As a broad variety of peptides is found in the secretions of amphibian skin combinatorial treatment of PGLa and magainin 2 was studied showing enhanced activity by a heterodimer formation. Thus production of mutually recognizing peptides seems to be an effective way in nature to increase selective membrane activity. Biophysical studies on membrane mimics demonstrated that PGLa can discriminate between different lipid species, preferentially interacting with negatively charged lipids, which are major components of bacterial but not mammalian cell membranes. This emphasizes the role of electrostatic interactions as a major determinant to trigger the affinity of antimicrobial peptides towards bacterial membranes. PGLa induced the formation of a quasi-interdigitated phase in phosphatidylglycerol bilayers below their chain melting transition, which is due to the creation of voids below the peptide being aligned parallel to the membrane surface. In the fluid phase of phosphatidylglycerol the peptide inserts perpendicularly into the bilayer above a threshold concentration, which results in a hydrophobic mismatch of the peptide length and bilayer core for lipids< or =C16. This mismatch is compensated by stretching of the acyl chains and in turn thickening of the bilayer demonstrating that membrane thinning cannot be taken generally as the hallmark of pore formation by antimicrobial peptides. Furthermore, PGLa was shown to affect membrane curvature strain of phosphatidylethanolamine, another main lipid component of bacterial membranes, where a cubic phase coexists with the fluid bilayer phase. Investigations on living Escherichia coli showed distinct changes in cell envelope morphology, when treated with the peptide. In a first stage loss of surface stiffness and consequently of topographic features was observed, followed in a second stage by permeabilization of the outer membrane and rupture of the inner (cytoplasmic) membrane supposedly by the mechanism(s) derived from model studies.

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Phosphatidylglycerol and cardiolipin represent the most abundant anionic phospholipid components of cytoplasmic bacterial membranes and thus are used as constituents for membrane mimetic systems. In this study, we have characterized the temperature dependent phase behaviour of the binary system dipalmitoyl-phosphatidylglycerol (DPPG) and tetramyristoyl-cardiolipin (TMCL) using microcalorimetry and X-ray scattering techniques. Both lipids exhibited a very similar main transition temperature (∼ 41 °C), showing a minimum (39.4 °C) for the binary mixtures at XDPPG = 0.8, and exhibited low-temperature phase transitions, which were abolished by incorporation of small amounts (≤ 10 mol%) of the other lipid component. Therefore, over a wide temperature and composition range a lamellar Lβ gel phase is the predominant structure below the chain melting transition, characterized by a relatively broad wide-angle peak for XDPPG ≤ 0.8. This observation suggests the existence of packing inconsistencies of the TMCL/DPPG hydrocarbon lattices in the gel phase, supported by the small average size of lipid clusters (∼ 50 lipids) within this composition range. The bilayer thickness for the lamellar-gel phase showed a monotonic increase (56 Å for TMCL to about 58 Å for XDPPG = 0.8 at 30 °C), which may be explained by different degrees of partial interdigitation of the acyl chains to compensate for the differences in the hydrocarbon lengths of DPPG and TMCL in the Lβ phase.

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