Phase separation and fluctuations in mixtures of a saturated and an unsaturated phospholipid.

We describe quantitatively the interactions in a mixture of a saturated and an unsaturated phospholipid, and their consequences to the phase behavior at macroscopic and microscopic levels. This type of lipid-lipid interaction is fundamental in determining the organization and physical behavior of biological membranes. Mixtures of dipalmitoylphosphatidylcholine (DPPC) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) are examined in detail by multiple experimental approaches (differential scanning calorimetry (DSC), fluorescence resonance energy transfer, and confocal fluorescence microscopy) in combination with Monte Carlo simulations in a lattice. The interactions between all possible pairs of lipid species and states are determined by matching the heat capacity calculated through Monte Carlo simulations to that measured experimentally by DSC. Only for one other lipid system, a mixture between two saturated phosphatidylcholines, is a similar quantitative description available. The interactions in the two systems and different representations used to model them are compared. Phase separation occurs in DPPC/POPC at about the center of the phase diagram mapped by DSC, but not at all compositions and temperatures in the coexistence region. Close to the extremes of composition, the phase behavior is best described by large fluctuations. At the heat capacity maxima in the mixtures, the domain size distributions change remarkably; large domains disappear and cooperative fluctuations increase.

What determines the activity of antimicrobial and cytolytic peptides in model membranes.

We previously proposed three hypotheses relating the mechanism of antimicrobial and cytolytic peptides in model membranes to the Gibbs free energies of binding and insertion into the membrane [Almeida, P. F., and Pokorny, A. (2009) Biochemistry 48, 8083-8093]. Two sets of peptides were designed to test those hypotheses, by mutating of the sequences of δ-lysin, cecropin A, and magainin 2. Peptide binding and activity were measured on phosphatidylcholine membranes. In the first set, the peptide charge was changed by mutating basic to acidic residues or vice versa, but the amino acid sequence was not altered much otherwise. The type of dye release changed from graded to all-or-none according to prediction. However, location of charged residues in the sequence with the correct spacing to form salt bridges failed to improve binding. In the second set, the charged and other key residues were kept in the same positions, whereas most of the sequence was significantly but conservatively simplified, maintaining the same hydrophobicity and amphipathicity. This set behaved completely different from predicted. The type of release, which was expected to be maintained, changed dramatically from all-or-none to graded in the mutants of cecropin and magainin. Finally, contrary to the hypotheses, the results indicate that the Gibbs energy of binding to the membrane, not the Gibbs energy of insertion, is the primary determinant of peptide activity.