Endogenous cell respiration was assayed in whole cells in the presence of galactose using a Clark-type polarographic oxygen electrode from Hansatech Instruments (Norfolk, UK) at 24°C as described (32 (link)).
Mitochondria were prepared from the different strains as described (6 (link)) and used for spectrophotometric assays performed at 24°C to measure KCN-sensitive COX activity, antimycin A-sensitive NADH cytochrome c reductase, and succinate cytochrome c reductase activities and oligomycin-sensitive ATP synthase activity, as described (32 (link)). Total mitochondrial cytochrome spectra were obtained as reported (32 (link)).
The abundance of OXPHOS complexes was analyzed by Blue Native polyacrylamide gel electrophoresis (BN-PAGE) using a linear 3–12% gradient gel (33 ).
Mitochondria were prepared from the different strains as described (6 (link)) and used for spectrophotometric assays performed at 24°C to measure KCN-sensitive COX activity, antimycin A-sensitive NADH cytochrome c reductase, and succinate cytochrome c reductase activities and oligomycin-sensitive ATP synthase activity, as described (32 (link)). Total mitochondrial cytochrome spectra were obtained as reported (32 (link)).
The abundance of OXPHOS complexes was analyzed by Blue Native polyacrylamide gel electrophoresis (BN-PAGE) using a linear 3–12% gradient gel (33 ).
