title: Interactions between protein 4.1 and band 3. An alternative binding site for an element of the membrane skeleton.
abstract: Protein 4.1 from human erythrocytes formed a complex with band 3 in inside-out erythrocyte membrane vesicles and with soluble peptides derived from the cytoplasmic domain of band 3. Protein 4.1 labeled metabolically with 32P bound saturably to vesicles depleted of endogenous protein 4.1. The soluble cytoplasmic domain of band 3 (43K) competitively displaced approximately 60% of bound 32P-protein 4.1 from reconstituted membrane vesicles. Pretreatment of vesicles with anti-43K similarly inhibited the rebinding of protein 4.1. In solution, 125I-43K formed a complex with protein 4.1 that saturated at 1:1 stoichiometry and migrated as a discrete band when analyzed by nondenaturing polyacrylamide gel electrophoresis. In rate-zonal sedimentation in isotonic salt solutions, protein 4.1 and 43K sedimented as a sharp peak at 4.4 S. In experiments aimed at exploring the role of the protein 4.1-band 3 interaction in the organization of the membrane skeleton, the effect of spectrin was investigated. Spectrin and protein 4.1 formed a complex which co-sedimented in sucrose gradients, but the addition of 43K to preformed spectrin-protein 4.1 complexes resulted in disruption of the complex and co-sedimentation of most of the protein 4.1 with 43K. These results suggest that protein 4.1 can associate with band 3 in the erythrocyte membrane and that this association may modulate the attachment of the membrane skeleton to the membrane.
mesh_terms: D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D001665:Binding Sites; D001667:Binding, Competitive; D001798:Blood Proteins / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins; D009479:Neuropeptides
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D001798:Blood Proteins; D003598:Cytoskeletal Proteins; D008565:Membrane Proteins; D009479:Neuropeptides; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; D005033:Ethylmaleimide