FIG. 7.

Preprotein binding to hTom22 and hTom20. (A) ³⁵S-labeled translation product (10 μl) was incubated for 30 min at 25°C with glutathione-agarose prebound with about 0.56 nmol of GST-(1–82)hTom22 or GST-(25–145)hTom20. After washing, GST derivatives were eluted with 15 mM reduced glutathione, and 40% of the eluted protein was subjected to SDS–10% PAGE and fluorography using a FUJIX BAS2000 analyzer as described in Materials and Methods. Five percent of input preproteins were put in the first lanes. R23A, R23A pOTC-GFP; R15/23/26A, R15/23/26A pOTC-GFP. (B) The radioactive preproteins eluted were quantified by image plate analysis. The binding was expressed as percentage of input precursors. Values are represented as means ± standard deviations of three independent experiments. (C) The binding assay was performed as described for panel A except that the indicated concentrations of KCl or Triton X-100 were added to binding and washing buffers. Binding was expressed as percentage of controls without KCl and Triton X-100.