Diffusion as a probe of the heterogeneity of antimicrobial peptide-membrane interactions

Abstract

Many antimicrobial peptides (AMPs) function by forming various oligomeric structures and/or pores upon binding to bacterial membranes. Because such peptide aggregates are capable of inducing membrane thinning and membrane permeabilization, we expected that AMP binding would also affect the diffusivity or mobility of the lipid molecules in the membrane. Herein, we show that measurements of the diffusion times of individual lipids through a confocal volume via fluorescence correlation spectroscopy (FCS) provide a sensitive means of probing the underlying AMP-membrane interactions. In particular, results obtained with two well-studied AMPs, magainin 2 and mastoparan X, and two model membranes indicate that this method is capable of revealing structural information, especially the heterogeneity of the peptide-membrane system, that is otherwise difficult to obtain using common ensemble methods. Moreover, because of the high sensitivity of FCS, this method allows examination of the effect of AMPs on the membrane structure at very low peptide/lipid ratios.

Figure 1

τ_D-distributions of TR-DHPE in GUVs composed of POPC and POPC/POPG, as indicated.

Figure 2

τ_D-distributions of TR-DHPE in POPC/POPG GUVs obtained at different bulk mag2 concentrations, as indicated.

Figure 3

Comparison of the τ_D-distributions of TR-DHPE in POPC/POPG GUVs obtained with and without the presence of mag2.

Figure 4

τ_D-distributions of TR-DHPE in POPC GUVs obtained at different bulk mag2 concentrations, as indicated.

Figure 5

τ_D-distributions of TR-DHPE in POPC/POPG GUVs obtained at different bulk mpX concentrations, as indicated.

Figure 6

τ_D-distributions of TR-DHPE in POPC GUVs obtained at different bulk mpX concentrations, as indicated.

Figure 7

Comparison of the τ_D-distributions of TR-DHPE in POPC GUVs obtained with and without the presence of mpX.