Surface exposure of phosphatidylserine is associated with the swelling and osmotically-induced fusion of human erythrocytes in the presence of Ca2+.
An assay for procoagulant activity has been used to investigate the Ca2(+)-dependent exposure of phosphatidylserine at the surface of human erythrocytes that were induced to swell and to fuse osmotically. Since the phosphatidylserine of human erythrocytes is located in the inner leaflet of the plasma membrane, it is inaccessible in intact cells which therefore had no procoagulant activity in an isotonic solution of sucrose. The procoagulant activity of erythrocytes incubated in increasingly hypotonic, sucrose solutions containing Mg2+ paralleled the percentage haemolysis, reflecting the accessibility of phosphatidylserine in an increasing number of lysed cells. However, cells in mildly hypotonic sucrose solutions containing Ca2+ had an abnormally high procoagulant activity indicating that phosphatidylserine was exposed in intact cells under these conditions. Erythrocytes that were subjected to continuous swelling at 37 degrees C, which was induced by entry of the permeant molecule poly(ethylene glycol) 400 (PEG 400) developed procoagulant activity in the presence of Ca2+ prior to extensive lysis. Cells treated in this way also fused. With Mg2+, PEG 400-treated erythrocytes lysed without fusing, and the development of procoagulant activity paralleled the rate of lysis. Erythrocytes incubated with ionophore A23187, subtilisin, and Ca2+ developed procoagulant activity (with less than 20% lysis), and they fused on subsequent exposure to a hypotonic medium. The procoagulant activity reached its maximum before fusion could be induced in the hypotonic medium. It is concluded that the entry of Ca2+ facilitates a translocation of phosphatidylserine to the outer leaflet of the erythrocyte plasma membrane that plays an important role in fusion protocols that involve cell swelling. It is also suggested that transbilayer movements of phosphatidylserine could be an important control factor in the cell biology of membrane fusion phenomena.
Department of Biochemistry and Chemistry, Royal Free Hospital School of Medicine, London U.K.
J M
JM
Baldwin
R
R
O'Reilly
M
M
Whitney
J A
JA
Lucy
D001692:Biological Transport; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology*; D004903:Erythrocyte Aggregation; D004910:Erythrocyte Membrane / Q000187:drug effects; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007038:Hypotonic Solutions; D008561:Membrane Fusion; D009997:Osmotic Pressure; D010718:Phosphatidylserines / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D013395:Sucrose
D007038:Hypotonic Solutions; D010718:Phosphatidylserines; D000001:Calcimycin; D011092:Polyethylene Glycols; D013395:Sucrose; D002118:Calcium