ABSTRACT
The study of the electrical parameters of asolectin bilayer lipid membranes in the presence of cytochrome c (cyt c) at various concentrations showed that an increase in the concentration of cyt c leads to an increase in the membrane conductance and the appearance of through pores. The studied membranes did not contain cardiolipin, which is commonly used in studying the effect of cyt c on membrane permeability. In the presence of cyt c, discrete current fluctuations were recorded. The occurrence of these fluctuations may be associated with the formation of through pores. The diameter of these pores was ~0.8 nm, which is smaller than the size of the cyt c globule (~3 nm). Measurements carried out at pH values from 6.4 to 8.4 showed that the concentration dependence of the membrane conductance increases with increasing pH. To assess the binding of cyt c to the bilayer, we measured the concentration and pH dependences of the difference in surface potentials induced by the unilateral addition of cyt c. The amount of bound cyt c at the same concentrations decreased with increasing pH, which did not correspond to the conductance trend. An analysis of conductance traces leads to the conclusion that an increase in the integral conductance of membranes is associated with an increase in the lifetime of pores. The formation of "long-lived" pores, of which the residence time in the open state is longer than in the closed state, was achieved at various combinations of pHs and cyt c concentrations: the higher the pH, the lower the concentration at which the long-lived pores appeared and, accordingly, a higher conductance was observed. The increase in conductance and the formation of transmembrane pores are not due to the electrostatic interaction between cyt c and the membrane. We hypothesize that an increase in pH leads to a weakening of hydrogen bonds between lipid heads, which allows cyt c molecules to penetrate into the membrane. This disrupts the order of the bilayer and leads to the occurrence of through pores.
ABSTRACT
We measured the conductance of bilayer lipid membranes of diphytanoylphosphatidylcholine induced by interaction with cubic magnetic nanoparticles (MNPs) of cobalt ferrite 12 and 27 nm in size and coated with a hydrophilic shell. The MNP coating is human serum albumin (HSA) or polyethylene glycol (PEG). The interaction of nanoparticles added to the bulk solution with the lipid bilayer causes the formation of metastable conductive pores, which, in turn, increases the integral conductance of the membranes. The increase in conductance with increasing MNP concentration was practically independent of the particle size. The dependence of the bilayer conductance on the concentration of PEG-coated MNPs was much weaker than that on the concentration with a shell of HSA. Analyzing the current traces, we believe that the conductive pores formed as a result of the interaction of nanoparticles with the membrane can change their size, remaining metastable. The form of multilevel current traces allows us to assume that there are several metastable pore states close in energy. The average radius of the putative cylindrical pores is in the range of 0.4-1.3 nm.
