title: Detection of the associated state of membrane proteins by polyacrylamide gradient gel electrophoresis with non-denaturing detergents. Application to band 3 protein from erythrocyte membranes.
abstract: Polyacrylamide gradient gel electrophoresis was carried out in micellar solutions of various detergents which differ in degree of potency to denature proteins. From the application of this method to band 3 protein from erythrocyte membranes, it was suggested that the procedure was useful in studying the molecular state of membrane proteins. The electrophoretic behaviors of human and bovine band 3 protein did not show any species specificity in either a denature state and a state resembling the native state. As well as in nonionic detergent solutions, the dimeric and tetrameric structures of bovine band 3 protein were preserved in sodium deoxycholate solution, in which protein complexes maintained in nonionic detergent solutions are frequently dissociated. Even in dodecyltrimethylammonium bromide solution, which is a denaturant for water-soluble proteins, part of the band 3 protein was still present as the oligomer. The results suggest that the oligomeric form of band 3 protein is the stable structure and that the dimer and tetramer possibly coexist in membranes.
mesh_terms: D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte; D001798:Blood Proteins / Q000032:analysis*; D002417:Cattle; D003840:Deoxycholic Acid; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004910:Erythrocyte Membrane / Q000032:analysis*; D004912:Erythrocytes / Q000032:analysis*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000097:blood*; D011489:Protein Denaturation; D000644:Quaternary Ammonium Compounds; D012967:Sodium Dodecyl Sulfate; D013045:Species Specificity; D013501:Surface-Active Agents
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D001798:Blood Proteins; D003902:Detergents; D008565:Membrane Proteins; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; D003840:Deoxycholic Acid; C013912:decyltrimethylammonium; D012967:Sodium Dodecyl Sulfate