The effect of oxidative stress on structural transitions of human erythrocyte ghost membranes.
Differential scanning microcalorimetry was used to study the effect of oxidative stress induced by cumene hydroperoxide (CHP) and Fe2+ on structural transitions of membranes of human erythrocyte ghosts. The CHP homolysis was shown to cause: (a) reduction of the intensity of all structural transitions with the disappearance of B1- and D-transitions; (b) decrease in the enthalpy of oxidized membrane denaturation; (c) negative slope of thermograms; (d) anomalous growth of heat absorption by membranes above 72 degreesC. All these changes occurred until the ratio Fe2+/CHP/membranes<0.02:0.05:1 was reached, i.e., prior to the moment of maximal level of TBA-RS in membrane ghosts. We interpret changes in the character of heat absorption by oxidized membranes as perturbations in the structural organization and interactions inside the spectrin-actin-protein 4.1 domains, the spectrin-protein 4.2 domain, as well as inside the domain of spectrin-ankyrin-cdB3 and the domain formed by the msdB3. These perturbations are associated mainly with the decrease in the concentration of native protein in the domains because of oxidative aggregation of proteins, as evidenced by SDS electrophoresis of oxidized membranes. Preincubation of membranes with tocopherol did not block the aggregation of proteins in electrophoresis and the decrease in the intensity of structural transitions, whereas it blocked completely the formation of TBA-RS, changes in the thermogram slope and the sharp rise in the heat absorption above 72 degreesC. This proves that these processes are determined by the thermotropic properties of the oxidized lipid bilayer of membranes and also provides evidence that the degradation of PUFA of phospholipids modifies both the structure of protein domains and the physical properties of the lipid bilayer of membranes.
Akoev
V R
VR
Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow Region 142292, Russian Federation. akoev@venus.iteb.serpukhov.su
Matveev
A V
AV
Belyaeva
T V
TV
Kim
Y A
YA
D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism; D002152:Calorimetry, Differential Scanning; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D006801:Humans; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D013696:Temperature; D014810:Vitamin E / Q000494:pharmacology
D001457:Anion Exchange Protein 1, Erythrocyte; D014810:Vitamin E