Cardiolipin Structure and Oxidation Are Affected by Ca2+ at the Interface of Lipid Bilayers

Abstract

Ca²⁺-overload contributes to the oxidation of mitochondrial membrane lipids and associated events such as the permeability transition pore (MPTP) opening. Numerous experimental studies about the Ca²⁺/cardiolipin (CL) interaction are reported in the literature, but there are few studies in conjunction with theoretical approaches based on <i>ab initio</i> calculations. In the present study, the lipid fraction of the inner mitochondrial membrane was modeled as POPC/CL large unilamellar vesicles (LUVs). POPC/CL and, comparatively, POPC, and CL LUVs were challenged by singlet molecular oxygen using the anionic porphyrin TPPS4 as a photosensitizer and by free radicals produced by Fe²⁺-citrate. Calcium ion favored both types of lipid oxidation in a lipid composition-dependent manner. In membranes containing predominantly or exclusively POPC, Ca²⁺ increased the oxidation at later reaction times while the oxidation of CL membranes was exacerbated at the early times of reaction. Considering that Ca²⁺ interaction affects the lipid structure and packing, density functional theory (DFT) calculations were applied to the Ca²⁺ association with totally and partially protonated and deprotonated CL, in the presence of water. The interaction of totally and partially protonated CL head groups with Ca²⁺ decreased the intramolecular P-P distance and increased the hydrophobic volume of the acyl chains. Consistently with the theoretically predicted effect of Ca²⁺ on CL, in the absence of pro-oxidants, giant unilamellar vesicles (GUVs) challenged by Ca²⁺ formed buds and many internal vesicles. Therefore, Ca²⁺ induces changes in CL packing and increases the susceptibility of CL to the oxidation promoted by free radicals and excited species.