Room temperature electron spin resonance of superoxide dismutase-loaded lipososmes and erythrocytes. A direct approach to the interaction of O2- with cells

Human erythrocytes were enriched with bovine superoxide dismutase by fusion with liposomes containing the entrapped enzyme. Liquid solution ESR of intact cells at room temperature was used to measure directly the increase in the superoxide dismutase content. From the spectral characteristics (g-value and hyperfine splitting tensor), the structural integrity of the Cu site of the enzyme was found to be unaffected by the liposome preparation procedure or the incubation with cells. Changes in the ESR signal size were used to test directly the interaction of superoxide with the enzyme entrapped in liposomes or delivered to erythrocytes. It was found that the liposome-entrapped enzyme does not react with externally generated O−2, but once delivered to red blood cells this reaction can take place. This is the first demonstration of O−2-scavenging activity by superoxide dismutase delivered into an intact cell structure and is therefore to be considered as strong evidence for activity of this enzyme under in vivo conditions.