40] Resolution and reconstitution of F0F1-ATPase in beef heart submitochondrial particles

Abstract

Publisher Summary his chapter surveys some current methods for the resolution and reconstitution of F 0 F l -ATPase with respect to F 1 , OSCP, and F 6 in beef heart submitochondrial particles. The oligomycin sensitivity conferring protein, OSCP and factor F6, are important with respect to their role in the interaction of F 1 with the membrane-bound, proton-translocating moiety (F 0 ) of the ATPase system, to yield an oligomycin-sensitive ATPase (F 0 F 1 -ATPase). The beef heart submitochondrial particles—vesiculated fragments of the mitochondrial inner membrane, with an inside-out orientation of the membrane surface—have been widely used in studies of the resolution and reconstitution of the enzyme system responsible for mitochondrial respiration and oxidative phosphorylation. The particles used in these experiments are prepared from beef heart mitochondria. These particles exhibit NADH and succinate oxidase activities and are non-phosphorylating, that is, they give rise to little or no ATP synthesis. Addition of oligomycin induces a high degree of respiratory control, and low concentrations of oligomycin can restore a limited capacity for oxidative phosphorylation. The particles, as prepared, exhibit relatively low ATPase activity because of the presence of the ATPase inhibitor protein.