Antimicrobial peptide alamethicin insertion into lipid bilayer: A QCM-D exploration

作者：Wang Kathleen F; Nagarajan Ramanathan; Camesano Terri A^*

来源：Colloids and Surfaces B: Biointerfaces , 2014, 116: 472-481.

DOI：10.1016/j.colsurfb.2014.01.036

• Antimicrobial peptide
• Lipid bilayer
• Alamethicin
• Phosphatidylcholine
• Quartz crystal microbalance
• Overtone

Alamethicin is a 20-amino-acid, alpha-helical antimicrobial peptide that is believed to kill bacteria through pore formation in the inner membranes. We used quartz crystal microbalance with dissipation monitoring (QCM-D) to explore the interactions of alamethicin with a supported lipid bilayer. Changes in frequency (Delta f) and dissipation (Delta D) measured at different overtones as a function of peptide concentration were used to infer peptide-induced changes in the mass and rigidity of the membrane as well as the orientation of the peptide in the bilayer. The measured Delta f were positive, corresponding to a net mass loss from the bilayer, with substantial mass losses at 5 mu M and 10 mu M alamethicin. The measured Delta f at various overtones were equal, indicating that the mass change in the membrane was homogeneous at all depths consistent with a vertical peptide insertion. Such an orientation coupled to the net mass loss was in agreement with cylindrical pore formation and the negligibly small Delta D suggested that the peptide walls of the pores stabilized the surrounding lipid organization. Dynamics of the interactions examined through Delta f vs. Delta D plots suggested that the peptides initially inserted into the membrane and caused disordering of the lipids. Subsequently, lipids were removed from the bilayer to create pores and alamethicin caused the remaining lipids to reorder and stabilize within the membrane. Based on model calculations, we concluded that the QCM-D data cannot confirm or rule out whether peptide clusters coexist with pores in the bilayer. We have also proposed a way to calculate the peptide-to-lipid ratio (P/L) in the bilayer from QCM-D data and found the calculated P/L as a function of the peptide concentration to be similar to the literature data for vesicle membranes.

• 出版日期2014-4-1