Oxidative red blood cell membrane injury in the pathophysiology of severe mouse beta-thalassemia.
In severe human beta-thalassemia, the pathophysiology relates to accumulation of excess alpha-globin chains at the membrane. One hypothesis is that membrane-associated alpha-globin by virtue of it's iron or hemichromes produces oxidation of adjacent membrane proteins. The availability of a mouse model of severe beta-thalassemia, as well as a transgenic (thalassemic-sickle) mouse that expresses 12% of human beta s-chain, has allowed us to study the effect of graded accumulation of alpha-chains at the red blood cell (RBC) membrane on the clinical status of the animal and on the material properties of its RBCs. Proteins from control, beta-thalassemic, and transgenic mouse RBC membranes were analyzed for evidence of oxidation, as measured by thiol-disulfide exchange chromatography, which detects intramolecular sulfhydryl oxidation. Ratios of oxidized globin to protein 7 were calculated and increased amounts were seen in thalassemic mice as compared with control mice and transgenic mice. Furthermore, there were increased amounts of thiol-free protein 4.1 in the thalassemic mice, compared with very small amounts in the control mice and intermediate amounts in the transgenic mice. Membrane mechanical stability as assessed by ektacytometry showed that the thalassemic mouse RBCs were markedly unstable. Transgenic mouse RBCs showed intermediate levels of membrane instability compared with the controls. We propose that this oxidized globin, in conjunction with oxidized protein 4.1, accounts (at least in part) for membrane instability. A 12% increase in beta s-globin chain synthesis (by decreasing excess globin available) confers considerable protection against both oxidative damage and the consequent membrane instability.
Advani
R
R
Department of Medicine (Hematology), Stanford University Medical School, CA 94305-5112.
Rubin
E
E
Mohandas
N
N
Schrier
S L
SL
D000818:Animals; D015153:Blotting, Western; D002845:Chromatography; D003598:Cytoskeletal Proteins; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D004907:Erythrocyte Deformability; D004910:Erythrocyte Membrane / Q000473:pathology*; D005914:Globins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009479:Neuropeptides; D010084:Oxidation-Reduction; D013438:Sulfhydryl Compounds / Q000097:blood; D013789:Thalassemia / Q000097:blood*
D003598:Cytoskeletal Proteins; D004220:Disulfides; D008565:Membrane Proteins; D009479:Neuropeptides; D013438:Sulfhydryl Compounds; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; D005914:Globins