FIGURE 5.

C9 binds to the ATPase domain of mortalin-ELISA. A, C9, C8, and C7 adsorbed onto microtiter plate wells were incubated with His-tagged recombinant full-length mortalin, SBD, or ATPase domain. The wells were washed, treated with anti-His antibody, and peroxidase-labeled second antibody, developed with OPD and analyzed in an ELISA reader. *, p < 0.005; **, p < 0.001 between ATPase domain and SBD. Error bars, S.D. B, microtiter plate wells were coated with C9 overnight at 4 °C. His-tagged mortalin or its domains were preincubated with C8 or C9 (0.25 μm, C8₁ and C9₁ or 0.5 μm, C8₂ and C9₂) for 1 h at 37 °C. The mixtures were then added to the C9-coated wells. Binding of His-tagged mortalin and its domains was quantified with anti-His antibody as in A. *, p < 0.05; **, p < 0.005; ***, p < 0.001 relative to control (− competitor).