FIG. 8.

The cross-linked AAC import intermediate is protected by the outer membrane in strain Δ70k mitochondria. In vitro import and cross-linking with radiolabeled AAC into strain Δ70k mitochondria were performed as in Fig. 7A, followed by treatment with 100 μg of trypsin per ml on ice for 30 min. After the addition of 200 μg of soybean trypsin inhibitor (Tryp) per ml, immunoprecipitation (IP) with protein A-Sepharose beads containing immobilized rabbit IgGs monospecific for Tim9p (α9) was performed as in Fig. 7A. S, standard (as defined in the legend to Fig. 6).