FIGURE 6.

The 47-kDa adduct is formed in the absence of subunit e. A, wild-type mitochondria (wt), ΔTIM11 mitochondria (mit.), and 1% (w/v) Triton X-100 ΔTIM11 mitochondrial extract (TX-100 ext.) were incubated with or without 2 mm CuCl₂ for 15 min at 30 °C. The reactions were stopped upon the addition of 10 mm EDTA and 10 mm NEM. The samples were solubilized in SDS, and aliquots (30 μg of protein) were analyzed by Western blot. B, CuCl₂-treated ΔTIM11 mitochondria were extracted with digitonin at the indicated digitonin-to-protein ratios in the presence of 10 mm NEM. Digitonin extracts were submitted to CN-PAGE. The slab gels were revealed by the in-gel ATPase activity procedure. C, fragment of gels corresponding to dimeric and monomeric forms of ATP synthase were cut and analyzed by Western blot as in the legend for Fig. 3. Blots were revealed with a polyclonal antibody raised against the yeast subunit 6 (su.6).