Interaction of horse heart cytochrome c with lipid bilayer membranes: effects on redox potentials.

Abstract

Cyclic voltammetry has been used to study the effects of interactions between horse cytochrome c and solid-supported planar lipid membranes, comprised of either egg phosphatidylcholine (PC) or PC plus 20 mol.% cardiolipin (CL), on the redox potential and the electrochemical electron transfer rate between the protein and a semiconductor electrode. Experiments were performed over a wide range of cytochrome c concentrations (0-440 microM) at low (20 mM) and medium (160 mM) ionic strengths. Three types of electrochemical behavior were observed, which varied as a function of the experimental conditions. At very low cytochrome c concentration (approximately 0.1 microM), and under conditions where electrostatic forces dominated the protein-lipid membrane interaction (i.e., low ionic strength with membranes containing CL), a redox potential (approximately 265 mV) and an electrochemical electron transfer rate constant (0.09 s[-1])were obtained which compare well with those measured in other laboratories using a variety of different chemical modifications of the working electrode. Two other electrochemical signals (not reported with chemically modified electrodes) were also observed to occur at higher cytochrome c concentrations with this membrane system, as well as with two other systems (membranes containing CL under medium ionic strength conditions, and PC only at low ionic strength). These involved positive shifts of the cytochrome c redox potential (by 40 and 60 mV) and large decreases in the electron transfer rate (to 0.03 and 0.003 s[-1]). The observations can be rationalized in terms of a structural model of the cytochrome c-membrane interaction, in which association involves both electrostatic and hydrophobic forces and results in varying degrees of insertion of the protein into the hydrophobic interior of the membrane.