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.
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.
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
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