Interaction of Arginine and Tryptophan-Rich Short Antimicrobial Peptides with Membrane Models: A Combined Fluorescence, Simulations, and Theoretical Approach.

The augmented increase in bacterial antimicrobial resistance necessitates the discovery of alternative antimicrobial molecules such as short antimicrobial peptides (AMPs) with antimicrobial activity and low cytotoxicity. While many such peptides have been studied, their selective affinity for bacterial versus mammalian membranes remains unclear. Here, we propose a complementary approach using state-of-the-art fluorescence experiments, molecular dynamics simulations, and theoretical techniques. The main goal of this approach is to unravel the energetics and molecular interactions of AMPs with different membrane models at the lipid-water interface. We use short Trp- and Arg-rich AMPs, pure phosphatidylcholine (PC), and an 85:15 mixture of PC with phosphatidylglycerol (PG) lipids for the mammalian and bacterial model membranes, respectively. First, we found that the electrostatic interaction of PG headgroups with Arg enhances the peptide interaction with mixed bilayers by 25-30%, leading to increased hydrogen bonding and stronger membrane adhesion. Second, the obtained Gibbs free energies revealed significantly distinct partitioning of the AMP at the interface for the two bilayers, suggesting a qualitatively different insertion method of cationic AMPs into each of the two membrane models. These results highlight the potential of our approach to unravel the membrane selectivity of an AMP in the context of AMP-based rational design of antibiotics.