Vesicle-to-cell protein transfer: insertion of band 3, the erythrocyte anion transporter, into lymphoid cells.
Band 3, the erythrocyte anion transporter, transfers spontaneously between human red cells and model membranes. During incubation of intact erythrocytes with sonicated dimyristoylphosphatidylcholine vesicles, the transporter inserts in functional form and native orientation into the liposome bilayer, with the cytoplasmic segment of the protein contacting the lumen of the vesicle [Newton, A. C., Cook, S. L., & Huestis, W. H. (1983) Biochemistry 22, 6110-6117; Huestis, W. H., & Newton, A. C. (1986) J. Biol. Chem. 261, 16274-16278]. When band 3-vesicle complexes are incubated with erythrocytes whose native band 3 has been inhibited irreversibly, reverse transfer of the protein restores anion transport capacity to the cells [Newton, A. C., Cook, S. L., & Huestis, W. H. (1983) Biochemistry 22, 6110-6117]. Here we report the vesicle-mediated transfer of band 3 to human peripheral blood lymphocytes and to cultured murine lymphoma cells (BL/VL3). Subsequent to incubation with protein-vesicle complexes, both lymphoid cell types exhibit a 2-4-fold increase in the rate of chloride uptake. This enhanced permeability is inhibited greater than or equal to 98% by the exofacial band 3 inhibitor 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid, consistent with right-side-out insertion of functional band 3 into the lymphoid cell membrane.
Newton
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Department of Chemistry, Stanford University, California 94305.
Huestis
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WH
D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D002460:Cell Line; D002712:Chlorides / Q000097:blood / Q000378:metabolism; D004134:Dimyristoylphosphatidylcholine; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008051:Lipid Bilayers; D008214:Lymphocytes / Q000378:metabolism*; D008223:Lymphoma / Q000378:metabolism; D051379:Mice; D008954:Models, Biological
D001457:Anion Exchange Protein 1, Erythrocyte; D002712:Chlorides; D008051:Lipid Bilayers; D004134:Dimyristoylphosphatidylcholine