Phlorizin- and 6-ketocholestanol-mediated antagonistic modulation of alamethicin activity in phospholipid planar membranes.

As a result of the interfacial chemical heterogenity, membrane-penetrating peptides will experience a dramatic variation in environmental polarity manifested via electrical interactions with the surface and dipole potential of membranes prone to modulate the membrane insertion and folding of different peptides and proteins. Herein we present evidence demonstrating that roughly a 30 mV, phlorizin-induced lowering of the magnitude of the dipole potential of a phosphatidyilcholine membrane leads to a 4-fold increase in the electrical activity of embedded alamethicin. The effect is voltage-independent, implying that the dipole potential affects the barrier of alamethicin adsorption to the membrane rather than the translocation of it across the hydrophobic core. Our interpretation points to an enhanced interfacial accumulation of alamethicin monomers on the cis side of the membrane caused by a lower value of the cis dipole potential, which will promote an elevated activity of alamethicin oligomers across the membrane. As expected for a modestly selective ion channel, the enhancing effect of such dipole potential changes on the electrical conductivity is limited (80 +/- 3 pS before and 100 +/- 2 pS after phlorizin addition to the membrane, for the first conductive state of the channel). Our study emphasizes the possibility that, by manipulating at will the sign of change and the magnitude of the interfacial dipole field, it is possible to modulate the extent of the membrane penetration of ion-channel-forming peptides and thereby provide deeper insights into mechanisms of protein-lipid and protein-protein interactions within membranes.