title: A procedure for membrane-protein reconstitution and the functional reconstitution of the anion transport system of the human-erythrocyte membrane.
abstract: A short procedure for the isolation of band-3 protein, the protein responsible for anion exchange in erythrocytes, in a reasonable degree of purity was developed. Using this protein preparation and a novel procedure for membrane-protein reconstitution, vesicles displaying the basic features of the anion-exchange system of the erythrocyte were obtained. The reconstitution procedure is based on slow direct removal of Triton X-100 from aqueous lipid/detergent solutions. According to the composition of the reconstitution medium, either small single-walled or large multi-walled vesicles are obtained. The procedure conserves protein properties well, as is revealed by the similarity of the rates of SO4(2-) exchange in erythrocytes and reconstituted vesicles when corrected for the relevant volumes. A number of functional features of the exchange system were studied and compared with those of the native membrane.
mesh_terms: D000838:Anions; D002352:Carrier Proteins / Q000097:blood*; D002463:Cell Membrane Permeability; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism / Q000648:ultrastructure; D004912:Erythrocytes / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D007474:Ion Exchange; D007700:Kinetics; D008565:Membrane Proteins / Q000097:blood*; D008854:Microscopy, Electron; D013431:Sulfates / Q000378:metabolism
chemical_list: D000838:Anions; D002352:Carrier Proteins; D008565:Membrane Proteins; D013431:Sulfates