Electrochemical analysis of the effect of Ca2+ on cardiolipin-cytochrome c interaction

Mitochondrial Ca2+ has been considered a trigger for the release of cytochrome c, which is a critical and early event in the induction of cell apoptosis, although the molecular mechanism underlying this effect is still not fully understood. Here we investigate the interaction between cytochrome c and cardiolipin and the effect of Ca2+ on this interaction using electrochemical methods. Experimental results revealed that modification of cardiolipin onto the surface of a pyrolytic graphite electrode could lead to a rapid direct electron transfer of cytochrome c through the electrostatic interaction between the protein and the cardiolipin. Addition of Ca2+ to the test solution containing cytochrome c could cause the decrease of the redox peaks of the protein, and the peaks could be recovered when Ca2+ was chelated by ethylenediaminetetraacetate. The cardiolipin-cytochrome c interaction and the Ca2+ effect were also investigated with the variation of the charges of lipids, buffer solutions, reaction time, and valencies of cations for comparison. (c) 2006 Elsevier B.V. All rights reserved.