FIGURE 7.
Assembly and oligomerization state of subunit h-depleted ATP synthase. A, mitochondria from RFY5-1 strain grown in the absence or in the presence of doxycycline (Dox) were solubilized at different digitonin/protein ratio. After centrifugation, the mitochondrial complexes were separated by CN-PAGE. The gel was incubated with ATP-Mg2+ and lead acetate to reveal the ATPase activity. For representation purposes, the figure is the negative of the original gel. B, samples from δh-mitochondria 2 g·g-1 digitonin extract were separated by CN-PAGE in duplicate: the 1st lane was revealed by ATPase activity (upper part of the B); the 2nd lane was separated in a second dimension by SDS-PAGE, analyzed by Western blotting, and revealed by anti-6 and anti-h polyclonal antibodies (lower part of B). (dim., dimer; mon., monomer).