Topo-optical studies of gradually disintegrated erythrocyte membrane derivates: different kinds of ghosts

Several kinds of ghosts from human erythrocytes (blood-group A1D) were investigated by the topo-optical Toluidine-Blue (TB) reaction. In comparison to intact cells, all ghosts demonstrated a decreased TB-anisotropy. These results reflect an altered glycocalyx structure of ghosts, especially conformational changes of the TB-binding N-terminal extracellular segments of the glycophorines. It was assumed that this structural glycocalyx alteration was caused by substantial losses of membrane skeleton components during the ghost preparation. Moreover, disturbed molecular interactions between the membrane skeleton and the glycocalyx may contribute to this effect. Therefore, the glycocalyx and the membrane structure of ghosts in general are significantly different from the membrane of the intact erythrocyte. The experiments show that the effects of the reconstitution procedures for ghost membranes are restricted to a reconstitution of a defined membrane function (i.e. dynamic barrier for monovalent cations) in a widely disturbed membrane structure.