1. Mitochondrial ribosomes remain associated with the inner membrane in the absence of the ribosome‐binding domain of Oxa1 and of Mba1.
Mitochondrial ribosomes remain associated with the inner membrane in the absence of the ribosome‐binding domain of Oxa1 and of Mba1. (A) Mitochondria from the strains indicated were incubated with or without puromycin and disintegrated by freeze thawing in 150 mM KCl, 5 mM EDTA, 20 mM Tris/HCl, pH 7.4. Then, the suspension was adjusted to 2 M sucrose and layers of 2 M sucrose, 1.5 M sucrose, 1 M sucrose in 60 mM KCl, 20 mM Tris/HCl, pH 7.4, were placed on top. After centrifugation at g for 16 h at 2°C, the gradient was fractionated. Proteins in the fractions were precipitated by the addition of 12% trichloroacetic acid and analyzed by Western blotting with antibodies against the proteins indicated. (B) Isolated wild‐type mitochondria were lysed in 0.5% dodecyl maltoside (DDM), 5 mM EDTA, 20 mM Tris/HCl, pH 7.4, for 15 min at 4°C and processed as in (A).
Martin Ott et al. EMBO J. 2006;25:
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