The coiled-coil domains of Mieap are critical for its interaction with BNIP3 and NIX.

<p>(A) The Mieap α and Mieap β mutant proteins. The adenoviral vectors expressing deletion mutants lacking the C-terminal region (α1–273 and β1–241) and the coiled-coil regions (ΔCC) of Mieap α and Mieap β proteins were prepared as indicated. (B–C) The mutants lacking coiled-coil regions of Mieap α and Mieap β proteins fail to bind NIX and BNIP3. HCT116 cells were transfected with plasmids expressing FLAG-NIX and FLAG-BNIP3 and infected with adenoviral vectors expressing the various Mieap α and Mieap β mutants. Cell lysates were isolated 2 days after transfection and infection and subjected to IP experiments. FLAG-NIX and FLAG-BNIP3 or the Mieap α and Mieap β mutants were precipitated using anti-FLAG (B) or anti-Mieap (C) antibodies, respectively. The precipitates were subjected to western blot analysis using anti-Mieap or anti-FLAG antibodies.