Removal of lipid from intact erythrocytes and ghosts by aqueous solutions and its relevance to membrane structure

Molar values for cholesterol, total phospholipid, and individual phospholipid classes of intact erythrocytes and their membranes (ghosts) washed with various aqueous solutions are presented. The data show that lipid can be washed from erythrocyte ghosts prepared rapidly from freshly drawn blood but that lipid is not removed from intact erythrocytes under the same conditions. Thus, it appears that the polar groups of lipids of intact cells are not exposed as they are in ghosts. In the preparation of hemoglobin-free ghosts, up to 25% cholesterol and phospholipid can be removed, while loss of ca. 50% cholesterol and phospholipid from ghosts can be achieved with aqueous solutions containing ethylenediamine tetraacetate. No significant loss of membrane protein was encountered even when almost half of the lipid had been removed from the ghosts. Phospholipid classes were removed to different extents with different wash solutions. Lipid loss from ghosts can be prevented, in part, by adding 0.5% albumin or calcium to wash solutions containing ethylenediamine tetraacetate. These findings contrast a report where insignificant lipid loss was noted in the preparation of hemoglobin-free human erythrocyte membranes, but agree with results reported for bovine red cell ghosts.