Protein-dependent lipid lateral phase separation as a mechanism of human erythrocyte ghost resealing.
The hypothesis of a correlation between a 10 degrees-20 degrees C lipid phase transition and the resealing process of human erythrocyte membrane has been investigated. The conditions required to reseal human erythrocyte ghosts have been studied by measuring the amount of fluorescein-labeled dextran (FD) that is trapped into the membrane. Temperature per se was sufficient to induce membrane resealing: (1) at 5 mM sodium phosphate, pH 7.8 (5P8), resealing began at 12 degrees C; (2) at salt concentrations above 8 mM sodium phosphate, it occurred at lower temperature; and (3) in isotonic saline was detected just above 5 degrees C. The removal of peripheral membrane proteins from unsealed membranes by chymotrypsin at 0 degree C in 5P8 was followed by membrane resealing. This seems to imply that the presence of proteins is necessary to maintain the membrane unsealed. Protein-induced lateral phase separation of lipids may be a reasonable mechanism for the observed phenomena. In fact, the permeability of phosphatidylserine-phosphatidylcholine mixed liposomes to FD is modified by lipid lateral phase separation induced by pH or poly-L-lysine. Electron spin resonance studies of membrane fluidity by a spin labeled stearic acid showed a fluidity break around 11 degrees C, which may be due to a gel-liquid phase transition. Fluidity changes are abolished by chymotrypsin treatment. It is suggested that a lateral phase separation is responsible for the permeability of open ghosts to FD. Accordingly, disruption of phase separation apparently produces membrane reconstitution. In this respect peripheral proteins and particularly the spectrin-actin network, may play a major role in membrane resealing.
D002463:Cell Membrane Permeability; D002918:Chymotrypsin / Q000494:pharmacology; D003911:Dextrans; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D005452:Fluoresceins; D006801:Humans; D066298:In Vitro Techniques; D008081:Liposomes; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000097:blood*; D008565:Membrane Proteins / Q000097:blood*; D009994:Osmolar Concentration; D013696:Temperature
D003911:Dextrans; D005452:Fluoresceins; D008081:Liposomes; D008563:Membrane Lipids; D008565:Membrane Proteins; D002918:Chymotrypsin