Radiation-induced Structural Changes in Membrane Proteins of Human Erythrocytes and Ghosts and the Relation to Cellular Morphology

Abstract

Isolated human erythrocytes and ghosts were irradiated with X-rays under different experimental conditions. The effect of the radiation treatment was examined with regard to the structure of membrane proteins as well as to the morphology of whole cells and ghosts. From sodium dodecyl sulphate/polyacrylamide gel electrophoresis it is concluded that spectrin (band 1 and 2) is the most radiosensitive of the membrane proteins examined. X-irradiation of cells and ghosts induced covalent cross-linking of a small fraction of membrane proteins. In the protein aggregates thus formed spectrin was found to be the major component. Molecular disulphide (-SS-) bridges seemed to account for part of the cross-links observed. Some nondisulphide cross-links were also found, especially when ghosts were irradiated. Significant amounts of spectrin aggregates were formed during post-irradiation incubation at 37°C but not at 4°C. In the intact cell a transformation in shape from discocyte to echinocyte accompanied the process of post-irradiation spectrin aggregation. The characteristics of both processes, such as their reversibility with adenosine, point to a metabolic involvement. It is shown that there is no causal relationship between the two phenomena observed. The possible cause of the post-irradiation effects and the parallelism with similar processes in nonirradiated metabolically depleted cells is discussed.