Cholesterol Affects the Pore Formation and the Membrane-Membrane Interaction Induced by an Antimicrobial Peptide, NK-2, in Phospholipid Vesicles.

Antimicrobial peptides are part of the innate immune response and show their antimicrobial activity by forming pores, followed by disintegration of the membrane. Cholesterol in the membrane can affect the pore formation process, as cholesterol is known to alter the permeability and elastic properties of the membrane. The present research systematically explores the role of cholesterol in modulating the interaction of the antimicrobial peptide NK-2 with phospholipid membranes, as well as the processes of pore formation induced by NK-2 within the membrane. Large unilamellar vesicles (LUVs) and giant unilamellar vesicles (GUVs) made from DOPC-DOPG and Egg PC with varying cholesterol concentrations have been studied using a variety of experimental techniques. The present study revealed that both the magnitude of zeta potential and surface charge density diminished as cholesterol concentrations increased at an intermediate NK-2 concentration. The proliferation of the size distributions of LUVs containing cholesterol when exposed to NK-2 indicates the occurrence of vesicle aggregation. The phase contrast micrographs of GUVs as well as the calcein release experiments on LUVs show evidence of pores. Notably, the incorporation of cholesterol into the membrane was found to have a significant effect on both the permeability of the membrane and the kinetics of the pore formation process. This biophysical research contributes essential knowledge regarding the role of cholesterol in influencing the antimicrobial efficacy of the membrane.