The role of phospholipid asymmetry in calcium-phosphate-induced fusion of human erythrocytes.
To elucidate the role of phospholipid asymmetry in calcium-phosphate-induced fusion of human erythrocytes, we examined the interaction of erythrocyte membranes with asymmetric and symmetric bilayer distributions of phospholipids. Fusion of human erythrocytes was monitored by light microscopy as well as spectrophotometrically by the octadecylrhodamine dequenching assay. Phospholipid translocation and distribution between the inner and the outer leaflet of intact red blood cells were determined with spin-labeled phosphatidylserine (PS), phosphatidylethanolamine (PE), and phosphatidylcholine (PC). Significant fusion of lipid-asymmetric red blood cells where PS and PE are predominantly oriented to the inner leaflet was only observed at Ca2+ concentrations greater than or equal to 10 mM (in the presence of 10 mM phosphate buffer) while fusion of lipid-symmetric erythrocyte membranes was established at greater than or equal to 1.5 mM Ca2+. The Ca2+ threshold of fusion of lipid-asymmetric red blood cells was significantly reduced (i) after exposure of PS to the outer layer but not after redistribution of PE alone, and (ii) upon incorporation of spin-labeled PS into the outer leaflet of red blood cells. Spin-labeled PE or PC did not affect fusion, suggesting that the serine headgroup is an important factor in calcium-phosphate-induced fusion.
Schewe
M
M
Institut für Biophysik, Humbolt-Universität Berlin, Germany.
Müller
P
P
Korte
T
T
Herrmann
A
A
D000001:Calcimycin / Q000494:pharmacology; D002130:Calcium Phosphates / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000502:physiology*; D004912:Erythrocytes / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D008051:Lipid Bilayers; D008561:Membrane Fusion / Q000187:drug effects*; D010743:Phospholipids / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013113:Spin Labels
D002130:Calcium Phosphates; D008051:Lipid Bilayers; D010743:Phospholipids; D013113:Spin Labels; D000001:Calcimycin