Anionic Lipids Confine Cytochrome c₂ to the Surface of Bioenergetic Membranes without Compromising Its Interaction with Redox Partners

Cytochrome c₂ (cyt. c₂) is a major element in electron transfer between redox proteins in bioenergetic membranes. While the interaction between cyt. c₂ and anionic lipids abundant in bioenergetic membranes has been reported, their effect on the shuttling activity of cyt. c₂ remains elusive. Here, the effect of anionic lipids on the interaction and binding of cyt. c₂ to the cytochrome bc₁ complex (bc₁) is investigated using a combination of molecular dynamics (MD) and Brownian dynamics (BD) simulations. MD is used to generate thermally accessible conformations of cyt. c₂ and membrane-embedded bc₁, which were subsequently used in multireplica BD simulations of diffusion of cyt. c₂ from solution to bc₁, in the presence of various lipids. We show that, counterintuitively, anionic lipids facilitate association of cyt. c₂ with bc₁ by localizing its diffusion to the membrane surface. The observed lipid-mediated bc₁ association is further enhanced by the oxidized state of cyt. c₂, in line with its physiological function. This lipid-mediated enhancement is salinity-dependent, and anionic lipids can disrupt cyt. c₂-bc₁ interaction at nonphysiological salt levels. Our data highlight the importance of the redox state of cyt. c₂, the lipid composition of the chromatophore membrane, and the salinity of the chromatophore in regulating the efficiency of the electron shuttling process mediated by cyt. c₂. The conclusions can be extrapolated to mitochondrial systems and processes, or any bioenergetic membrane, given the structural similarity between cyt. c₂ and bc₁ and their mitochondrial counterparts.