Evaluation of cytochrome c affinity to anionic phospholipids by means of surface plasmon resonance.

We attempted to evaluate the affinity of the anionic phospholipids to cytochrome c by means of surface plasmon resonance (SPR) technique and to correlate it with the cytochrome c active site alterations and peroxidase activity. Our experiments showed a strong interdependence between the phospholipid fatty acid saturation degree, the active site structure alterations and peroxidase activity of the cytochrome c phospholipid complex. Cytochrome c peroxidase activity and Trp59 fluorescence increase in the sequence of phosphatidyl choline (PC)-->phosphatidylserine (PS)-->cardiolipin (CL)-->phosphatidic acid (PA). The association constant (K(a)) increased in the sequence PC-->PA-->PS-->CL. The SPR spectroscopy data shows that K(a) is independent of lipid saturation degree, but correlates with phospholipid negative charge value.