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. 1999 Mar 30;96(7):3634–3639. doi: 10.1073/pnas.96.7.3634

Figure 4.

Figure 4

Chase of pSu9-DHFR from stage B but not from stage A depends on the presence of the IMS domain of Tom22. (A) pSu9-DHFR was bound to CCCP-treated mitochondria prepared from MNMS-MAS17 (WT) or from MNMS-MAS17Δ120–152 (ΔC) at 0°C (lanes 1–8) or at 30°C (lanes 9–16). The samples were divided into four aliquots, which were diluted 5-fold with MSC buffer containing different concentrations of KCl to result in final KCl concentrations of 10 mM KCl (lanes 1, 2, 9, and 10), 50 mM KCl (lanes 3, 4, 11, and 12), 100 mM KCl (lanes 5, 6, 13, and 14), or 150 mM KCl (lanes 7, 8, 15, and 16) and incubated for 5 min at 0°C. The samples were divided into halves and the mitochondria were reisolated by centrifugation. One aliquot was resuspended with binding buffer and kept on ice (odd-numbered lanes). The other aliquot was resuspended with chase buffer and incubated for 10 min at 30°C (even-numbered lanes). p and m, precursor and mature forms of pSu9-DHFR, respectively. (B Upper) Quantification of the bound precursor form (odd-numbered lanes of A). (Lower) Chased mature-sized form (even-numbered lanes of A). The amount of the bound precursor form at 10 mM KCl at 0 or 30°C is set to 100%.