Gramicidin A Channel Formation Induces Local Lipid Redistribution I: Experiment and Simulation

Integral membrane protein function can be modulated by the host bilayer. Because biological membranes are diverse and nonuniform, we explore the consequences of lipid diversity using gramicidin A channels embedded in phosphatidylcholine (PC) bilayers composed of equimolar mixtures of di-oleoyl-PC and di-erucoyl-PC (dC_18:1+dC_22:1, respectively), di-palmitoleoyl-PC and di-nervonoyl-PC (dC_16:1+dC_24:1, respectively), and di-eicosenoyl-PC (pure dC_20:1), all of which have the same average bilayer chain length. Single-channel lifetime experiments, molecular dynamics simulations, and a simple lipid compression model are used in tandem to gain insight into lipid redistribution around the channel, which partially alleviates the bilayer deformation energy associated with channel formation. The average single-channel lifetimes in the two-component bilayers (95 ± 10 ms for dC_18:1+dC_22:1 and 195 ± 20 ms for dC_16:1+dC_24:1) were increased relative to the single-component dC_20:1 control bilayer (65 ± 10 ms), implying lipid redistribution. Using a theoretical treatment of thickness-dependent changes in channel lifetimes, the effective local enrichment of lipids around the channel was estimated to be 58 ± 4% dC_18:1 and 66 ± 2% dC_16:1 in the dC_18:1+dC_22:1 and dC_16:1+dC_24:1 bilayers, respectively. 3.5-μs molecular dynamics simulations show 66 ± 2% dC_16:1 in the first lipid shell around the channel in the dC_16:1+dC_24:1 bilayer, but no significant redistribution (50 ± 4% dC_18:1) in the dC_18:1+dC_22:1 bilayer; these simulated values are within the 95% confidence intervals of the experimental averages. The strong preference for the better matching lipid (dC_16:1) near the channel in the dC_16:1+dC_24:1 mixture and lesser redistribution in the dC_18:1+dC_22:1 mixture can be explained by the energetic cost associated with compressing the lipids to match the channel's hydrophobic length.