Kinetics of Channel Formation in Bilayer Lipid Membranes (BLMs) and Tethered BLMs: Monazomycin and Melittin

The kinetics of channel formation by the polyene-like antibiotic monazomycin, both in a bilayer lipid membrane (BLM) and in a tethered BLM (tBLM), and by the peptide melittin in a tBLM, is investigated. Stepping the applied potential from a value at which channels are not formed to one at which they are formed yields current vs time curves that are sigmoidal on a BLM, while they show a maximum on a tBLM; in the latter case, sigmoidal curves are obtained by plotting the charge against time. These curves are interpreted on the basis of a general kinetic model, which accounts for the potential-dependent penetration of adsorbed monomeric molecules into the lipid bilayer, followed by their aggregation with channel formation by a mechanism of nucleation and growth. In the case of monazomycin, which is present in the solution in the form of relatively hydrophilic clusters and is adsorbed as such on top of the lipid bilayer, penetration into the bilayer following a potential jump is assumed to be preceded by a potential-independent disaggregation of the adsorbed clusters into adsorbed monomers

Kinetics of Channel Formation in Bilayer Lipid Membranes (BLMs) and Tethered BLMs: Monazomycin and Melittin

The kinetics of channel formation by the polyene-like antibiotic monazomycin, both in a bilayer lipid membrane (BLM) and in a tethered BLM (tBLM), and by the peptide melittin in a tBLM, is investigated. Stepping the applied potential from a value at which channels are not formed to one at which they are formed yields current vs time curves that are sigmoidal on a BLM, while they show a maximum on a tBLM; in the latter case, sigmoidal curves are obtained by plotting the charge against time. These curves are interpreted on the basis of a general kinetic model, which accounts for the potential-dependent penetration of adsorbed monomeric molecules into the lipid bilayer, followed by their aggregation with channel formation by a mechanism of nucleation and growth. In the case of monazomycin, which is present in the solution in the form of relatively hydrophilic clusters and is adsorbed as such on top of the lipid bilayer, penetration into the bilayer following a potential jump is assumed to be preceded by a potential-independent disaggregation of the adsorbed clusters into adsorbed monomers

Gramicidin Conducting Dimers in Lipid Bilayers Are Stabilized by Single-File Ionic Flux along Them

Gramicidin D was incorporated in a biomimetic membrane consisting of a lipid bilayer tethered to a mercury electrode via a hydrophilic spacer, and its behavior was investigated in aqueous 0.1 M KCl by potential-step chronocoulometry and electrochemical impedance spectroscopy. The impedance spectra, recorded from 0.1 to 1 × 105 Hz over a potential range of 0.7 V, were fitted to a series of RC meshes, which were related to the different substructural elements of the biomimetic membrane. These impedance spectra were compared with those obtained by incorporating valinomycin, under otherwise identical conditions. The potential dependence of the stationary currents reported on bilayer lipid membranes by Bamberg and Läuger (Bamberg, E.; Läuger, P. J. Membrane Biol. 1973, 11, 177−194) as well as those extracted from potential-step chronocoulometric measurements was interpreted by relating the increase in gramicidin dimerization to a progressive increase in single-file K+ flux along the dimeric channels. An analogous approach was adopted in explaining the difference between the impedance spectra obtained with gramicidin D and those obtained with valinomycin. It is concluded that gramicidin has a low tendency to form dimers in the absence of ionic flux

Impedance Spectroscopy of OmpF Porin Reconstituted into a Mercury-Supported Lipid Bilayer

The channel-forming protein OmpF porin was incorporated in a biomimetic membrane consisting of a lipid bilayer tethered to a mercury electrode via a thiolipid, and it was investigated in aqueous KCl by electrochemical impedance spectroscopy. The impedance spectra, recorded from 1 × 10-2 to 1 × 105 Hz over a potential range of 0.7 V, were fitted to an equivalent circuit consisting of four RC meshes. The dependence of the resulting circuit elements upon the applied potential was interpreted on the basis of a general approximate approach based on a model of the electrified interphase and on the kinetics of the translocation of potassium and chloride ions across the lipid bilayer, assisted by the OmpF porin

Interaction of Mixed-Ligand Monolayer-Protected Au₁₄₄ Clusters with Biomimetic Membranes as a Function of the Transmembrane Potential

Understanding the interaction of nanoparticles with cell membranes is a high-priority research area for possible biomedical applications. We describe our findings concerning the interaction of Au144 monolayer-protected clusters (MPCs) with biomimetic membranes and their permeabilizing effect as a function of the transmembrane potential. We synthesized Au144(SCH2CH2Ph)60 and modified the capping monolayer with 8-mercaptooctanoic acid (Au144OctA) or thiolated trichogin (Au144TCG), a channel-forming peptide. The interactions of these MPCs with mercury-supported lipid mono- and bilayers were studied with a combination of electrochemical techniques specifically sensitive to changes in the properties of biomimetic membranes and/or charge-transfer phenomena. Permeabilization effects were evaluated through the influence of MPC uptake on the reduction of cadmium­(II) ions. The nature and properties of the Au144 capping molecules play a crucial role in controlling how MPCs interact with membranes. The native MPC causes a small effect, whereas both Au144OctA and Au144TCG interact significantly with the lipid monolayer and show electroactivity. Whereas Au144OctA penetrates the membrane, Au144TCG pierces the membrane with its peptide appendage while remaining outside of it. Both clusters promote Cd2+ reduction but with apparently different mechanisms. Because of the different way that they interact with the membrane, Au144OctA is more effective in Cd2+ reduction when interacting with the lipid bilayer and Au144TCG performs particularly well when piercing the lipid monolayer

Potassium Ion Transport by Valinomycin across a Hg-Supported Lipid Bilayer

A biomimetic membrane consisting of a lipid bilayer tethered to a mercury electrode via a hydrophilic spacer was investigated in aqueous KCl by potential-step chronocoulometry and electrochemical impedance spectroscopy, both in the absence and in the presence of the ionophore valinomycin. Impedance spectra, recorded from 1 × 10-2 to 1 × 105 Hz over a potential range of 0.8 V, are satisfactorily fitted to a series of four RC meshes, which are straightforwardly related to the different substructural elements of the biomimetic membrane. The frequency-independent resistances and conductances of both the lipid bilayer and the hydrophilic spacer show a maximum when plotted against the applied potential. This behavior is interpreted on the basis of a general approximate approach that applies the concepts of impedance spectroscopy to a model of the electrified interphase and to the kinetics of potassium ion transport assisted by valinomycin across the lipid bilayer