FIG 5.

All the TM regions of Tim17 are essential for Tim23 interaction. (A to E) Purified mitochondria from the wild type and tim17 mutants (TM1 to -4) were subjected to heat shock at 37°C for 30 min to induce phenotype. The mitochondria were lysed with digitonin-containing buffer, and the lysates were subjected to co-IP using anti-FLAG antibody. Samples were separated on SDS-PAGE gels and subjected to Western blot analysis using the indicated specific antibodies to detect the different components of presequence translocase. Twenty-five percent of the total sample served as loading control (Input) against 100% of immunoprecipitated product. The amounts of different immunoprecipitated proteins were quantified by densitometry using ImageJ software.