Abnormal membrane protein methylation and merocyanine 540 fluorescence in sickle erythrocyte membranes.
Sickle cell erythrocytes exhibit reduced carboxyl methylation of membrane proteins compared to normal erythrocytes. This altered methylation in sickle membrane proteins is also observable when extracted membranes, both intact and alkali treated, were used as substrates for the homologous protein methylase II (S-adenosylmethionine:protein-carboxyl O-methyltransferase, EC. 2.1.1.24). However, when glycophorin A, one of the major methyl acceptors in both membranes, was extracted by lithium diiodosalicylate and used as the methyl acceptor, the proteins from both membranes were methylated equally, suggesting an involvement of membrane structure in membrane-bound protein methylation. Merocyanine 540 (MC-540), a fluorescent probe, was used to determine if the membranes differed in organization. Incubation of both normal and sickle erythrocytes membranes with MC-540 produced a marked increase in extrinsic fluorescence, reflecting a relatively nonpolar environment for the dye bound to the membranes. The fluorescence from sickle cell ghosts was only 87% as intense as that from normal ghosts, while the actual amount of MC-540 associated with sickle cell membranes was only 62% of normal. These data suggest that differences exist in the distribution of surface charges on these plasma membranes. These results are consistent with the hypothesis that abnormal levels of membrane protein methylation observed in sickle erythrocytes may be a result of abnormal membrane organization characteristic to sickle cell anemia.
D000755:Anemia, Sickle Cell / Q000097:blood*; D001798:Blood Proteins / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004913:Erythrocytes, Abnormal / Q000378:metabolism*; D005456:Fluorescent Dyes; D006021:Glycophorins / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000097:blood*; D008745:Methylation; D011744:Pyrimidinones; D013050:Spectrometry, Fluorescence
D001798:Blood Proteins; D005456:Fluorescent Dyes; D006021:Glycophorins; D008565:Membrane Proteins; D011744:Pyrimidinones; C003954:merocyanine dye