Differential sensitivity of canine cardiac sarcolemmal and microsomal enzymes to inhibition by free radical-induced lipid peroxidation

Abstract

Sarcolemmal and microsomal membranes prepared from adult canine cardiac myocytes (sarcolemmal Na+,K+-ATPase = 71.8 μmol/mg per hr and microsomal rotenone-insensitive NADH cytochrome c reductase = 114 μmol/mg per hr) were each preincubated at 37°C in the presence of a free radical generating system consisting of dihydroxyfumarate and Fe+++-ADP; loss of the Na+,K+-ATPase and reductase activities, as well as the associated increases in lipid peroxidation, measured by malondialdehyde formation, were temporally correlated in both systems. The ATPase was inhibited 70% when the malondialdehyde was 71 nmol/mg protein at 20 minutes and 90% when malondialdehyde was 138 nmol/mg protein at 90 minutes. Inhibition of reductase activity occurred more gradually, displaying a 27% loss of activity when malondialdehyde reached 34 nmol/mg protein at 20 minutes and 60% with a malondialdehyde value of 67 nmol/mg protein at 90 minutes. The greater susceptibility of the sarcolemma to free radical-induced membrane damage may be due to the higher content of unsaturated fatty acids in this membrane, compared to microsomes.