Separation of the lipid bilayer from the membrane skeleton during discocyte-echinocyte transformation of human erythrocyte ghosts.
The membrane skeleton, a protein lattice at the internal side of the red cell membrane, is principally composed of spectrin, actin and proteins 4.1 and 4.9. We have examined negatively stained red cell ghosts and demonstrated, on an ultrastructural level, a separation of the lipid bilayer from the membrane skeleton during echinocytic transformation. The electron micrographs of discoidal red cell ghosts suspended in hypotonic buffer revealed a filamentous reticulum that uniformly laminated the entire submembrane region. transformation of the discoidal ghosts into echinocytic form, as induced by incubation in isotonic buffer, resulted in a disruption of skeletal continuity underlying the surface contour of the membrane spicule. The submembrane reticulum extended into the base and the neck of the spiny processes of the crenated ghosts but was absent at the tip of these projections. In addition, membrane vesicles without a submembrane reticulum were detected either attached to the tips of the spicules or released into the supernatant from the echinocytic ghosts. Protein analysis revealed that the released vesicles were enriched in bands 3, 4.1 and 7 and contained very little of the membrane skeletal proteins, spectrin and actin. The data indicate that during echinocyte formation, parts of the lipid bilayer physically separate from the membrane skeleton, leading to a formation of skeleton-poor lipid vesicles.
Department of Biomedical Research, St. Elizabeth's Hospital, Boston, MA 02135.
S C
SC
Liu
L H
LH
Derick
M A
MA
Duquette
J
J
Palek
D000199:Actins / Q000032:analysis; D004910:Erythrocyte Membrane / Q000032:analysis / Q000648:ultrastructure*; D006651:Histocytochemistry; D006801:Humans; D008051:Lipid Bilayers / Q000032:analysis*; D008565:Membrane Proteins / Q000032:analysis; D008854:Microscopy, Electron; D008954:Models, Biological; D013049:Spectrin / Q000032:analysis
D000199:Actins; D008051:Lipid Bilayers; D008565:Membrane Proteins; D013049:Spectrin