Identification of the protein 4.1 binding interface on glycophorin C and p55, a homologue of the Drosophila discs-large tumor suppressor protein.
Protein 4.1 is the prototype of a family of proteins that include ezrin, talin, brain tumor suppressor merlin, and tyrosine phosphatases. All members of the protein 4.1 superfamily share a highly conserved N-terminal 30-kDa domain whose biological function is poorly understood. It is believed that the attachment of the cytoskeleton to the membrane may be mediated via this 30-kDa domain, a function that requires formation of multiprotein complexes at the plasma membrane. In this investigation, synthetically tagged peptides and bacterially expressed proteins were used to map the protein 4.1 binding site on human erythroid glycophorin C, a transmembrane glycoprotein, and on human erythroid p55, a palmitoylated peripheral membrane phosphoprotein. The results show that the 30-kDa domain of protein 4.1 binds to a 12-amino acid segment within the cytoplasmic domain of glycophorin C and to a positively charged, 39-amino acid motif in p55. Sequences similar to this charged motif are conserved in other members of the p55 superfamily, including the Drosophila discs-large tumor suppressor protein. Our data provide new insights into how protein 4.1, glycophorin C, p55, and their non-erythroid homologues, interact with the cytoskeleton to exert their physiological effects.
Marfatia
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SM
Department of Biomedical Research, St. Elizabeth's Medical Center, Tufts University School of Medicine, Boston, Massachusetts 02135.
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Chishti
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003598:Cytoskeletal Proteins; D017931:DNA Primers; D004330:Drosophila / Q000378:metabolism; D029721:Drosophila Proteins; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006021:Glycophorins / Q000378:metabolism*; D051528:Guanylate Kinases; D006801:Humans; D007301:Insect Hormones / Q000235:genetics; D008565:Membrane Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009479:Neuropeptides; D009703:Nucleoside-Phosphate Kinase / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D025521:Tumor Suppressor Proteins
D003598:Cytoskeletal Proteins; D017931:DNA Primers; D029721:Drosophila Proteins; D006021:Glycophorins; D007301:Insect Hormones; D008565:Membrane Proteins; D009479:Neuropeptides; D025521:Tumor Suppressor Proteins; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1; C070350:dlg1 protein, Drosophila; D009703:Nucleoside-Phosphate Kinase; D051528:Guanylate Kinases