Research Paper Volume 5, Issue 4 pp 234—269

Figure 6. Under CR conditions, the atg32Δ mutation eliminates the non-respiratory protein supercomplex 5 in the IMM and alters compositions of other protein supercomplexes recovered by 1-D BN-PAGE.

WT and atg32Δ strains were cultured in the nutrient-rich YP medium initially containing 0.2% glucose. Mitochondria were purified from WT and atg32Δ cells recovered on day 4 of culturing. Digitonin-solubilized protein supercomplexes from the inner membrane of mitochondria purified from WT (A) or atg32Δ (B) cells were separated by first-dimension blue native PAGE (1-D BN-PAGE) and then resolved by second-dimension tricine-SDS-PAGE (2-D SDS-PAGE). Following silver staining, the separated by 2-D SDS-PAGE proteins were identified by mass spectrometry. Arrows in A mark the non-respiratory protein supercomplex 5 lacking in the IMM of atg32Δ cells as well as individual proteins or respiratory protein complexes dissociated from other protein supercomplexes in the IMM of these mutant cells.