Figure 5.

PMH1/PMH2 are bound in RNA-dependent high-molecular-mass complexes. DDM-solubilized total mitochondrial protein from cell suspension culture was separated in the first dimension by BN-PAGE followed by urea SDS-PAGE in the second dimension. A to E, Coomassie-stained gel (A); immunodetection analysis of DDM-solubilized protein with PMH1/PMH2 antibodies (B); as B after separation of RNase A-treated mitochondrial lysates (C); immunodetection analysis with PMH1/PMH2 antibody after cold treatment of the suspension culture for 18 h (D); and solubilization in the presence of 10 mm MgCl₂ (E). Sizes in the first dimension are estimated from complexes I + III₂ (1,500 kD), I (1,000 kD), V (580 kD), III₂ (480 kD), formate dehydrogenase (FDH; 200 kD), and aconitase (100 kD; Eubel et al., 2005).