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. Author manuscript; available in PMC: 2008 Sep 15.
Published in final edited form as: FEBS J. 2007 Dec 6;275(1):44–58. doi: 10.1111/j.1742-4658.2007.06172.x

Fig. 3. Human eIF5A mutant proteins as substrates for deoxyhypusine synthase and deoxyhypusine hydroxylase in vitro.

Fig. 3

Human recombinant eIF5A proteins were expressed in E. coli, BL21(DE3) and cell lysates were used as substrates for DHS and DOHH in a combined assay. Coomassie-Blue staining of BL21(DE3) lysates expressing human mutant proteins (A, top panel) and fluorogram of the SDS gel of DHS/DOHH reaction mixture showing labeling of several mutant proteins (A, bottom panel). Portions of DHS/DOHH reaction mixtures were analyzed for radioactive deoxyhypusine and hypusine content in the products by ion exchange chromatographic separation (B) as described previously (49). The experiments were repeated two to three times with virtually the same results: a typical experiment is shown.