FIG. 1.

Association of CHC with Tom1L1. (A) Modular structure of Tom1L1 wild type and of the mutants used in this study. The VHS and GAT homology domains, the linker region, and the C terminus (C) are indicated. Src binding sites (i.e., PP₄₂₁LP and Tyr457), the mutations of these Src binding sites (i.e., A₄₂₀ALA and Phe457 in Tom1L1YPP), and the mutation of the CHC binding site A₄₀₁AQPSVA (in ΔL/L401A) are indicated. (B) Association of CHC with Tom1L1 in vitro. HeLa cell lysates were incubated with the indicated GST fusion proteins or control GST beads, and the presence of CHC in the pull-down assays was revealed by Western blotting (wb) with a specific antibody. Input (15% of the cell lysate) was loaded as a positive control. αCHC, anti-CHC. (C) Association of CHC with Tom1L1 in NIH 3T3 cells. The CHC level associated with Tom1L1 in NIH 3T3 cells was assessed by Western blotting with anti-CHC antibody after immunoprecipitation of Tom1L1 with control (immunoglobulin G [IgG]) or anti-Tom1L1.3 (αTom1L1.3) antibody as shown. The level of immunoprecipitated Tom1L1 is also shown.