ACBD5 interacts with VAPB. (A) Schematic overview of VAPB and ACBD5 domain structure. Mutations in acyl-CoA binding and FFAT-like motifs are indicated. (B) Immunoprecipitation (IP) of GFP-ACBD5 and Myc-VAPB after coexpression in COS-7 cells. GFP was used as control. Samples were immunoprecipitated (GFP-Trap) and immunoblotted using Myc-GFP antibodies. (C) In vitro binding assay using GST-VAPB and MBP fusions of ACBD5 (AcB, mutations in the acyl-CoA binding motif; FFAT, mutations in FFAT motif) expressed in E. coli. GST served as control. Samples were immunoprecipitated using GST-Trap and immunoblotted using MBP-GST antibodies. (D) Model of ACBD5–VAPB interaction.