Two-dimensional gel electrophoresis of membrane proteins using anionic and cationic detergents. Application to the study of mitochondrial F0-F1-ATPase.
Polyacrylamide gel electrophoresis in the presence of a cationic detergent, tetradecyltrimethylammonium bromide (TDAB) has been compared to electrophoresis in the presence of an anionic detergent, sodium dodecyl sulfate (SDS). Although, in both systems, the peptides generally migrated as a function of their molecular weight, the TDAB electrophoresis permitted us to obtain a much better resolution of several peptides of the mitochondrial F0-F1-ATPase, especially for the alpha and beta subunits and for the oligomycin sensitivity conferring protein (OSCP). The differences between the two electrophoretic profiles have been used to devise a new technique of two-dimensional electrophoresis using successively anionic and cationic detergents. This method could be very useful in the case of membrane proteins, which are generally soluble only in the presence of powerful ionic detergents. It has been particularly successful in resolving the small peptides of the F0-F1-ATPase which were difficult to differentiate by other techniques in one- or two-dimensional polyacrylamide gel electrophoresis.
D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D002352:Carrier Proteins; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000032:analysis*; D008929:Mitochondria, Heart / Q000201:enzymology*; D025261:Mitochondrial Proton-Translocating ATPases; D008970:Molecular Weight; D006180:Proton-Translocating ATPases / Q000032:analysis*; D000644:Quaternary Ammonium Compounds; D012967:Sodium Dodecyl Sulfate; D013552:Swine; D050337:Trimethyl Ammonium Compounds
D002352:Carrier Proteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D000644:Quaternary Ammonium Compounds; D050337:Trimethyl Ammonium Compounds; D012967:Sodium Dodecyl Sulfate; D000251:Adenosine Triphosphatases; D025261:Mitochondrial Proton-Translocating ATPases; D006180:Proton-Translocating ATPases; C020439:oligomycin sensitivity-conferring protein; C009410:tetradecyltrimethylammonium