Figure 4.

Specific interaction of the Tom22 precursor with the isolated TOB complex. TOB complex was isolated from OMVs on a Ni-NTA matrix using TX-100. It was incubated while bound to the matrix with radioactive precursors of the indicated proteins (bars in dark gray). As a control and reference, mock isolation without OMVs was performed and the matrix was treated the same way with radioactive precursor proteins (bars in light gray). After washing, the bound precursor proteins were eluted and visualized by autoradiography. Binding in the mock control to the Ni-NTA matrix was set to 1. The amounts of precursor bound to the TOB complex immobilized on the Ni-NTA matrix were related to the mock control level. Error bars: standard deviation of five experiments. Radioactive precursor proteins: Tom40, porin, Su9-DHFR (matrix-targeting sequence of ATPase subunit 9 fused to mouse dihydrofolate reductase), Tom22. Control lysate (no plasmid was added to the transcription–translation reaction).