Defective binding of spectrin to ankyrin in a kindred with recessively inherited hereditary elliptocytosis.
The interaction of spectrin with spectrin-depleted inside-out membrane vesicles was studied in a kindred with an atypical variant of hereditary elliptocytosis inherited in a recessive manner. The probands are characterized by prominent elliptocytosis, decreased erythrocyte thermal stability, an altered limited tryptic peptide pattern of spectrin digested at low ionic strength, and defective spectrin dimer-dimer association. The parents are normal. The spectrin/band 3 ratio determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of isolated membranes of the probands was decreased to approximately 70% of control values, and total erythrocyte spectrin content in one proband was also decreased on SDS-PAGE. When a monospecific antispectrin antibody was used, a faintly labeled fragment of molecular weight approximately 28,000 was detected on immunoblots of whole cell lysates of one proband and a control, but could not account for the decreased erythrocyte spectrin content of the proband on SDS-PAGE. Binding and competitive inhibition studies revealed an alteration in the spectrin-ankyrin interaction due to an abnormality of spectrin in the probands. No defect was found in the mother; the father's spectrin showed decreased binding affinity, although it was not so severe as in the probands. Separation of bound and unbound spectrin dimers from one proband and subsequent conversion to tetramers showed that the self-association defect was detectable only on the bound subpopulation of her spectrin. These findings demonstrate a hitherto undescribed functional abnormality of spectrin in this kindred which could result in decreased stability of the membrane skeleton and contribute to the elliptocytic shape of these erythrocytes.
D000328:Adult; D017487:Ankyrins; D002675:Child, Preschool; D004591:Electrophoresis, Polyacrylamide Gel; D004612:Elliptocytosis, Hereditary / Q000097:blood* / Q000235:genetics; D004910:Erythrocyte Membrane / Q000032:analysis; D005260:Female; D005808:Genes, Recessive; D006801:Humans; D007525:Isoelectric Focusing; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D013049:Spectrin / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D017487:Ankyrins; D008565:Membrane Proteins; D013049:Spectrin