Abstract

Antimicrobial agents (AMAs) often exert their effect by forming pores in the bacterial cell membrane, a process fundamentally governed by the lipid composition of the membrane. This study investigates the **mechanisms by which lipid environment regulates the pore-forming activity of AMAs**, a critical determinant of their efficacy and selectivity. Using the well-established **planar lipid bilayer (PLB) technique**, we reconstituted model membranes with varying lipid types and concentrations to systematically probe the AMA-membrane interaction. Our primary objective was to elucidate how specific lipid properties, such as charge, headgroup structure, and acyl chain saturation, modulate the kinetics of pore insertion, stability, and conductance. Key findings reveal that negatively charged lipids significantly enhance AMA binding and pore formation, leading to increased membrane permeability. Conversely, high cholesterol content was found to stabilize the bilayer, thereby reducing the frequency and duration of pore events. These results provide crucial molecular-level insights into the lipid-mediated control of AMA function, which can inform the rational design of new antimicrobial peptides and small molecules with improved therapeutic profiles and reduced off-target effects on host cell membranes.