Fig. 3.

TimA and TimB do not associate in a membrane complex. (A) Inner membrane vesicles (IM) solubilized with DDM were subject to immunoprecipitation assays with pre-immune serum (−) or antiserum recognizing TimA (A) or TimB (B). The immuno-depleted supernatants and the precipitates were analyzed by SDS/PAGE and immunoblotting with the sera as shown. (B) Immunoprecipitations from inner membrane vesicles (1 mg total protein) using either pre-immune serum (pre-) or immune serum raised to TimA were analyzed by SDS/PAGE and Coomassie staining, with the identity of the precipitated proteins determined by MS. (C) Inner membrane vesicles were solubilized with the detergent DDM and analyzed by blue native PAGE and immunoblotting for TimA and TimB. The Coomassie blue–stained gel (stain) is also shown.