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Fig. 7. Inhibition of protein import into both rat liver and yeast mitochondria by high ionic strength. (A) AAC and Su9-DHFR were synthesized in reticulocyte lysate in the presence of ³⁵S-methionine. Isolated rat liver mitochondria were added in the presence of increasing concentrations of KCl as indicated. The samples were incubated at 25°C for 20 minutes, cooled-down to 0°C and treated with proteinase K. The mitochondria were isolated again and analyzed by SDS-PAGE and fluorography. The relative amounts of imported protein were calculated, the values determined for the 80 mM KCl sample were set to 100% (control). (B) Import of pCIC and mCIC, following the same procedure as in A. (C) Precursor (pPiC) and mature phosphate carrier lacking the presequence (mPiC) were synthesized in reticulocyte lysate in the presence of ³⁵S-methionine. Isolated yeast mitochondria (wild-type) were added in the presence of increasing concentrations of KCl as indicated. The samples were then treated as described in (A). (D) pPiC and mPiC were imported into isolated yeast mitochondria as described in C. Standard samples of the reticulocyte lysate were included in the SDS-PAGE to determine the ratio of imported PiC vs the total amount of PiC added to each sample.

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