Figure 9.

Assembly of TIM23 complexes is altered in absence of Mmp37p. Mitochondria isolated from wild-type and mmp37Δ strains were solubilized with digitonin (1% wt/vol), clarified by centrifugation and either directly analyzed by BN-PAGE (A), or incubated with additional digitonin (final concentration 2% wt/vol), before the BN-PAGE step (B). After separation of solubilized complexes by BN-PAGE, the protein complexes were resolved by a second dimension of SDS-PAGE and were subsequently analyzed by Western blotting with anti-Tim23p, anti-Pam18p/Tim14p, anti-Pam16p/Tim16p, and anti-Mmp37p, as indicated.