Cytoskeleton-membrane connections in the human erythrocyte membrane: band 4.1 binds to tetrameric band 3 protein.
Band 4.1 provides, besides ankyrin, the main linkage between the erythrocyte membrane and its cytoskeleton. Its predominant binding sites in the membrane are located on the glycophorins. However, the cytoplasmic domain of band 3 can also bind band 4.1. We have studied which of the different band 3 oligomers observed (monomers, dimers, tetramers) can act as band 4.1 binding sites, by equilibrium sedimentation experiments on mixtures of purified band 3 and dye-labelled band 4.1 in solutions of a nonionic detergent. At low molar ratios of band 4.1 and band 3, the sedimentation equilibrium distributions obtained could all be perfectly fitted assuming that only two dye-labelled particles were present: uncomplexed band 4.1 and a complex formed between one band 4.1 molecule and one band 3 tetramer. The presence of small amounts of complexes containing band 3 monomers or dimers could not be completely ruled out but is unlikely. On the other hand, stabilized band 3 dimers effectively bound band 4.1. At higher molar band 4.1/band 3 ratio, the band 3 tetramer apparently could bind up to at least four band 4.1 molecules. The band 4.1/band 3 tetramer complex was found to be unstable. The results described, together with those reported previously, point at a prominent role of tetrameric band 3 in ligand binding.
von Rückmann
B
B
Institut für Biophysik, J.W. Goethe-Universität, Frankfurt am Main, Germany.
Jöns
T
T
Dölle
F
F
Drenckhahn
D
D
Schubert
D
D
D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D001665:Binding Sites; D003598:Cytoskeletal Proteins; D003599:Cytoskeleton / Q000378:metabolism*; D003902:Detergents; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D009479:Neuropeptides; D012996:Solutions
D001457:Anion Exchange Protein 1, Erythrocyte; D003598:Cytoskeletal Proteins; D003902:Detergents; D008565:Membrane Proteins; D009479:Neuropeptides; D012996:Solutions; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1