Fig. 1.
Expression of eIF5A, DHS and DOHH in BL21(DE3)pLysS cells. E.coli cells transformed with the pST39 empty vector (lane 1), pST39/eIF5A (lane 2), pST39/eIF5A/DHS (lane 3) or pST39/eIF5A/DHS/DOHH (lane 4) were induced with 1 mM IPTG for 4 h in the presence or absence of [3H]spermidine (5 μCi/ml). The protein expression was measured after SDS-PAGE by Coomassie-Blue staining (A) or by western blotting (B). Hypusine modification was measured by ion exchange chromatographic separation of radioactive deoxyhypusine or hypusine in the acid hydrolysate of proteins (C) and the total radioactivity in hypusine (lane 4) or deoxyhypusine peak (lane 3) in each sample is shown (D).