Phospholipid composition and organization in model beta-thalassemic erythrocytes.
The membrane phospholipid organization in human red blood cells (RBC) is rigidly maintained by a complex system of enzymes. However, several elements of this system are sensitive to oxidative damage. An important component in the destruction of beta-thalassemic RBC is the generation of reactive oxygen species and the release of redox-active iron by the unpaired alpha-hemoglobin chains. Consequently, we hypothesized that the presence of this oxidative stress to the RBC membrane could lead to alterations in membrane lipid organization. Model beta thalassemic RBC, prepared by the introduction of excess alpha-globin in the cell, have previously been shown to exhibit structural and functional changes almost identical to those observed in beta-thalassemic cells. After 24 hr at 37 degrees C, the model beta thalassemic cells exhibited a significant loss of deformability, as measured by ektacytometric analysis, indicative of extensive membrane damage. However, a normal steadystate distribution of endogenous phospholipids was found, as evidenced by the accessibility of membrane phospholipids to hydrolysis by phospholipases. Similarly, the kinetics of transbilayer movement of spin-labeled phosphatidylserine (PS) and phosphatidylethanolamine (PE) in all samples was in the normal range and was not affected by the presence of excess alpha-globin chains. In contrast, a faster rate of spin-labeled phosphatidylcholine (PC) transbilayer movement was observed in these cells. While control RBC exhibited a complete loss of their initial (2 mol%) lysophosphatidylcholine (LPC) levels following 24 hr of incubation at 37 degrees C, 1.5 mol% LPC was still present in model beta-thalassemic cells, suggesting an altered phospholipid molecular species turnover, possibly as a result of an increased repair of oxidatively damaged phospholipids.
Kuypers
F A
FA
Children's Hospital Oakland Research Institute, California 94609, USA.
Schott
M A
MA
Scott
M D
MD
D000255:Adenosine Triphosphate / Q000097:blood; D004907:Erythrocyte Deformability; D004910:Erythrocyte Membrane / Q000737:chemistry*; D005914:Globins / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008563:Membrane Lipids / Q000032:analysis*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010743:Phospholipids / Q000032:analysis*; D013113:Spin Labels; D017086:beta-Thalassemia
D008563:Membrane Lipids; D010743:Phospholipids; D013113:Spin Labels; D000255:Adenosine Triphosphate; D005914:Globins