The membrane lateral pressure-perturbing capacity of parabens and their effects on the mechanosensitive channel directly correlate with hydrophobicity

Lipid bilayers provide a natural anisotropic environment for membrane proteins and can serve as apolar reservoirs for lipid-derived second messengers or lipophilic drugs. Partitioning of lipophilic agents changes the lateral pressure distribution in the bilayer, affecting integral proteins. p-Hydroxybenzoic acid esters (parabens) are amphipathic compounds widely used as food and cosmetics preservatives, but the mechanisms of their broad antibacterial action are unknown. Here we describe effects of ethyl, propyl, and butyl parabens on the gating of the bacterial mechanosensitive channel of small conductance (MscS) and compare them with the surface activity and lateral pressure changes measured in lipid monolayers in the presence of these substances. Near the bilayer-monolayer equivalence pressure of 35 mN/m, ethyl, propyl, or butyl paraben present in the subphase at 1 mM increased the surface pressure of the monolayer by 5, 12.5, or 20%, respectively. No spontaneous activation of MscS channels was observed in patch-clamp experiments with parabens added from either the cytoplasmic or periplasmic side. Increasing concentrations of parabens on the cytoplasmic side of excised patches shifted activation curves of MscS toward higher tensions. A good correlation between the pressure increases in monolayers and shifts in activation midpoints in patch-clamp experiments suggested that the more hydrophobic parabens partition more strongly into the lipid and exert larger effects on channel gating through changes in lateral pressure. We show that cytoplasmically presented ethyl or butyl parabens both hasten the process of desensitization of MscS and influence inactivation differently. The higher rate of desensitization is likely due to increased lateral pressure in the cytoplasmic leaflet surrounding the gate. Neither of the parabens strongly affects the rate of recovery and does not seem to penetrate the TM2-TM3 interhelical clefts in MscS. We conclude that the bacterial mechanosensitive channel MscS provides a sensitive readout of lateral membrane pressure exerted by amphipathic molecules but may not be the primary target for the parabens in their antimicrobial activity.