Lipid peroxidation both inhibits Ca2+ATPase and increases Ca2+ permeability of endoplasmic reticulum membrane

Abstract

Incubation of reticular membranes with Fe2+-EDTA and H2O2 plus Fe2+-EDTA at 37°C for 30 min. led to the loss of membrane's efficiency to sequester Ca2+ to 21.8% and 3.6% of control values, respectively. The incubation of microsomes with Fe2+-EDTA and H2O2 plus Fe2+-EDTA also caused decrease of Ca2+-ATPase activity; to 44.9% and 44.4% (measured under the same conditions as Ca2+-uptake) or to 79.6% and 62.1% (uncoupled from Ca2+ transport by detergent). In addition, incubation of membranes with Fe2+-EDTA and H2O2 plus Fe2+-EDTA at 37°C for 30 min. led to the increase of Ca2+ permeability to 125.1% and 124.2%, respectively. Preincubation of membranes with membrane-soluble antioxidants (U-74500A, U-83836E, t-butyl hydroxytoluene and stobadine) protected the reticular membranes against depression of Ca2+ uptake values and Ca2+-ATPase inhibition in a dose and an antioxidant nature dependent manner. Our results indicate that both processes, Ca2+-ATPase inhibition and increase of endoplasmic reticulum membrane Ca2+ permeability, participate in the lipid peroxidation induced loss of membranes efficiency to sequester Ca2+.