Expression of recombinant Vhs and GST-eIF4H. (A) Coomassie blue-stained gel of recombinant proteins bound to glutathione-Sepharose and eluted with 10 mM glutathione. Proteins are from E. coli expressing GST-eIF4H (GST/4H) (lanes b to e) and wild-type (WT) Vhs (lane b), D194N (lane c), or D215N (lane d). Material in lane e is from bacteria expressing GST-eIF4H and no Vhs. Vhs is indicated by a closed circle to the right of lanes b to d, and GST-eIF4H is indicated by an arrow. (B) Material that eluted from glutathione-Sepharose was applied to a column of HiTrap SP Sepharose and eluted with a gradient of 0 to 1 M salt. Fractions were analyzed for Vhs by Western blotting. 4H, eIF4H. (C) Coomassie blue-stained gel of HiTrap SP Sepharose fractions 10 to 13 from cells cells expressing wild-type Vhs and GST-eIF4H. Vhs is indicated by a closed circle, and GST-eIF4H is indicated by an arrow.