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In vitro reconstitution of the first step of diphthamide biosynthesis with 14C-SAM. A) In vitro reconstitution of PhDph2 activity using dithionite. B) In vitro reconstitution of yeast Dph1-3 activity. Lane 2 is protein ladder. All the reactions in lanes 1, 3 and 4 contained EF2 and Dph1-Dph2. The presence of other reagents is indicated below each lane. In both A and B, the left panels show the Coomassie blue-stained gel; the right panels show the autoradiography.
