Extraction of erythrocyte membrane proteins by sulfhydryl inhibitors.
Human red cell membrane proteins were extracted by incubation of the ghost with hypotonic phosphate buffer (pH 7.4), N-ethylmaleimide and p-hydroxy-mercuribenzoate. In paroxysmal nocturnal hemoglobinuria (PNH), hereditary spherocytosis (HS) and hereditary elliptocytosis, the amount of proteins extracted by these procedures was significantly less than the amount extractable from the ghost of normal and aplastic anemia red cells. Polypeptide patterns of red cell membranes in these hematological disorders were essentially similar to those of normal ghosts. Analysis of the supernatant by SDS polyacrylamide gel electrophoresis revealed that this reduction was mainly due to the reduced amount of peripheral proteins extracted. The extraction of peripheral proteins by sulfhydryl reagents was accompanied by shape changes resulting in the formation of membrane vesicles, suggesting an important role of peripheral proteins in the maintenance of ghost shape. It is also suggested that qualitative abnormalities of peripheral proteins such as altered reactivity to sulfhydryl reagents and/or strong binding to the membrane are present in PNH, HS and hereditary elliptocytosis red cells.
D000743:Anemia, Hemolytic / Q000097:blood*; D004612:Elliptocytosis, Hereditary / Q000097:blood; D004910:Erythrocyte Membrane / Q000032:analysis / Q000473:pathology; D004912:Erythrocytes / Q000032:analysis; D005033:Ethylmaleimide; D006456:Hemoglobinuria / Q000097:blood; D006461:Hemolysis; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D013103:Spherocytosis, Hereditary / Q000097:blood; D013439:Sulfhydryl Reagents
D008565:Membrane Proteins; D013439:Sulfhydryl Reagents; D005033:Ethylmaleimide