Comparison of spectrin isolated from erythroid and non-erythroid sources.
Spectrin from erythrocytes and two other tissues (brain and intestine) were isolated from two distant species, pig and chicken; some structural and functional properties were compared. A quantitative antibody inhibition assay was used to determine that antibodies to mammalian red cell spectrin cross-react very poorly, if at all, with their non-erythroid (brain) counterpart and similarly antibodies to pig brain spectrin (fodrin) cross-react very weakly with erythroid spectrin. By contrast, antibodies which were directed against the 240000-Mr subunit of avian fodrin were completely inhibited with avian spectrin and vice versa. To analyze the structural relatedness of these molecules further we compared the chymotryptic iodinated peptide maps generated from each individual subunit. Consistent with the antibody results, we find little (less than 10%) homology between peptides derived from mammalian fodrin and spectrin, but complete homology (100%) of the peptides derived from the 240000-Mr subunits of chicken fodrin, spectrin and another related molecule from intestine, TW260/240. Whereas the peptide maps of fodrin (brain spectrin) revealed striking similarity between divergent species, suggesting a high degree of structural conservation, the peptide maps of erythrocyte spectrin was highly variable between species, indicating that it has diverged considerably in mammalian evolution. In addition we have compared a functional activity of mammalian spectrins, the ability to bind calmodulin, using two different assays. Both results show that, whereas fodrin-calmodulin interaction can be readily demonstrated, the binding to mammalian erythroid spectrin is negligible. This suggests that the high-affinity calmodulin site present on fodrin has been lost from spectrin in mammalian evolution.
D000818:Animals; D000906:Antibodies / Q000032:analysis; D001923:Brain Chemistry; D002147:Calmodulin / Q000378:metabolism; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology; D002645:Chickens; D004912:Erythrocytes / Q000032:analysis*; D007422:Intestines / Q000032:analysis*; D046911:Macromolecular Substances; D008840:Microfilament Proteins / Q000032:analysis / Q000276:immunology; D008970:Molecular Weight; D013049:Spectrin / Q000276:immunology / Q000302:isolation & purification*; D013552:Swine
D000906:Antibodies; D002147:Calmodulin; D002352:Carrier Proteins; D046911:Macromolecular Substances; D008840:Microfilament Proteins; D013049:Spectrin