FIG. 3.
Association of the head domain of vimentin with PMT. The lysates of E. coli expressing the head (H; amino acids 1 to 94), rod (R; amino acids 95 to 406), and tail (T; amino acids 407 to 465) domains of vimentin in the GST-tagged forms were subjected to the ligand overlay assay with PMT840-1285. Arrows indicate the predicted position of each domain. (A) The expression of the fusion proteins was estimated by Western blot analysis with anti-GST polyclonal antibody. The amount of each fragment was estimated from the intensity of the blotted protein band. (B) Ligand overlay assay. An equal amount of each fragment was applied to each lane. Note that the head domain at the predicted position was reacted with PMT840-1285, whereas the rod and the tail domains were not. The bands that appeared at the position below 25 kDa should be nonspecifically reactive proteins, because they were observed even in the lysate from naive E. coli. The positions of the molecular mass standards are shown on the left.