Modulation of Antimicrobial Peptide–Membrane Interactions by Lysyl-Phosphatidylglycerol in Staphylococcus aureus: An FTIR Spectroscopy Study

Changes in membrane lipid composition constitute a key bacterial resistance mechanism. In Staphylococcus aureus, phosphatidylglycerol undergoes lysine modification to form lysyl-phosphatidylglycerol, a cationic lipid that reduces the net negative surface charge and thereby enhances resistance to cationic antimicrobial peptides. In this study, we examined the influence of lysyl-PG on the membrane activity of three antimicrobial peptides with distinct physicochemical characteristics: LL-37, F5W Magainin II, and NA-CATH:ATRA-1-ATRA-1. Model membranes composed of phosphatidylglycerol and cardiolipin were supplemented with increasing molar fractions of lysyl-phosphatidylglycerol, and peptide–membrane interactions were characterized using Fourier-transform infrared spectroscopy. Membrane fluidity was evaluated through shifts in the symmetric methylene stretching bands, while changes in interfacial polarity were assessed via the carbonyl and phosphate asymmetric stretching bands. LL-37 induced pronounced disruption of anionic bilayers, an effect progressively attenuated by lysyl-phosphatidylglycerol, particularly within the hydrophobic core. F5W Magainin perturbed both hydrophobic and interfacial regions across a broader range of lysyl-phosphatidylglycerol concentrations, whereas NA-CATH:ATRA-1-ATRA-1 primarily targeted interfacial domains, with minimal disruption of acyl chain order. Increasing lysyl-PG content modulated the extent of bilayer disorder and dehydration at the hydrophobic–hydrophilic interface, with each peptide exhibiting a distinct interaction profile. Collectively, these findings provide mechanistic insights into lysyl-PG-mediated modulation of peptide activity and highlight the role of lipid remodeling as a bacterial defense strategy.