FIGURE 6.

C9 binds to the ATPase domain of mortalin-sucrose density gradient. A, C9 (1 μg) was mixed in an Eppendorf tube with recombinant His-tagged mortalin, SBD, or ATPase domain (2 μg) for 1 h at 37 °C. The samples were layered on top of 13-ml 10–30% sucrose density gradients and were sedimented for 18 h at 40,000 rpm at 4 °C. Fractions (300 μl) were collected from the gradient top and analyzed by dot blotting with anti-His antibody. B, density of each scanned dot was quantified with ImageJ software. Plotted dot density of mortalin or mortalin domains in fractions 1–16 is shown. A.U., arbitrary units.