Biophysical characterization of genistein-membrane interaction and its correlation with biological effect on cells - the case of EYPC liposomes and human erythrocyte membranes
Bożena Pawlikowska-Pawlęga, Lucjan E. Misiak, Anna Jarosz-Wilkołazka, Barbara Zarzyka, Roman Paduch, Antoni Gawron, Wieslaw I. Gruszecki
With application of EPR and 1H NMR techniques genistein interaction with liposomes formed with egg yolk lecithin and with erythrocyte membranes was assessed. The present study addressed the problem of genistein localization and its effects on lipid membrane fluidity and protein conformation. The range of microscopic techniques was employed to study genistein effects on HeLa cells and human erythrocytes. Moreover, DPPH bioassay, superoxide anion radicals test and enzymatic measurements were performed in HeLa cells subjected to genistein. The gathered results from both EPR and NMR techniques indicated strong ordering effect of genistein on the motional freedom of lipids in the head group region and the adjacent hydrophobic zone in liposomal as well as in red blood cells membranes. EPR study of human ghost showed also the changes in the erythrocyte membrane protein conformation. The membrane effects of genistein were correlated with the changes in internal membranes arrangement of HeLa cells as it was noticed using transmission electron microscopic and fluorescent techniques. Scanning electron and light microscopy methods showed that one of the aftermaths of genistein incorporation into membranes was creation of echinocytic form of the red blood cells with reduced diameter. Genistein improved redox status of HeLa cells treated with H2O2 by lowering radicals’ level.
In conclusion, the capacity of genistein to incorporate, affect membrane organization and to change its biophysical properties is correlated with the changes inside cells.