Thermal properties of young red blood cells are indicative of an age-dependent regulation of membrane-skeleton interaction.

Abstract

The effects of red blood cell (RBC) age on membrane thermal properties have been investigated by using a 16-nitroxide stearic acid spin probe. We detected in unfractionated and most dense cells (2% fraction of circulating cells) a thermal transition at 40 degrees C that in young cells (1% fraction) was lowered at 33-35 degrees C. Spectrin seems to be directly involved in the transition detected in both young and unfractionated cells, as showed by the disappearance of the breaks after low salt extraction of spectrin. A further indication for a role of spectrin in this transition comes from its characteristic thermal unfolding above 40 degrees C. However, young cells did not show changes either in the thermal unfolding of spectrin or in the distribution of spectrin dimer, tetramer, and high oligomeric forms. These data rule out that spectrin of young RBC is modified in its thermal properties and indicate that young cells may have a different spectrin-membrane interaction. Treatment of unfractionated ghosts with an antibody specific for a fragment of the 10K domain of protein 4.1, which is fully competent for the spectrin-actin binding, produced an evident lowering of the transition temperature. The same antibody did not affect the thermal transition of young ghosts. Our results suggest that spectrin-membrane interactions may be regulated during RBC lifespan.