title: Isolation of the mitochondrial F1-F0 adenosine triphosphatase by Sepharose-hexylammonium chromatography: properties and reconstitution in liposomes.
abstract: Lauryl dimethylamino oxide, a zwitterionic detergent, was employed to solubilize the H+ ATPase from beef heart mitochondria. A simple preparation procedure has been devised to obtain F1-F0 based on a method described to purify F1 ATPase (M. Tuena de Gómez-Puyou and A. Gómez-Puyou, 1977, Arch. Biochem. Biophys. 182, 82-86) which consists of the selective adsorption of F1 to Sepharose-hexylammonium beads. The preparation showed approximately 18 bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis; 5 correspond to F1 subunits and the rest probably to the stalk and hydrophobic sector F0. The binding of [14C]dicyclohexylcarbodiimide to a low-molecular-weight component of this preparation was demonstrated. The F1-F0 complex was reconstituted into phospholipid vesicles which displayed ATP-Pi exchange and ATP-dependent 9-aminoacridine fluorescence quenching, both sensitive to proton channel inhibitors.
mesh_terms: D000251:Adenosine Triphosphatases / Q000302:isolation & purification*; D000585:Aminacrine; D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D003902:Detergents; D004024:Dicyclohexylcarbodiimide; D004591:Electrophoresis, Polyacrylamide Gel; D008081:Liposomes / Q000032:analysis; D008565:Membrane Proteins / Q000302:isolation & purification; D008929:Mitochondria, Heart / Q000201:enzymology*; D006180:Proton-Translocating ATPases; D012995:Solubility; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling
chemical_list: D003902:Detergents; D008081:Liposomes; D008565:Membrane Proteins; D004024:Dicyclohexylcarbodiimide; D000585:Aminacrine; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases