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. 2014 Apr 16;136(17):6179–6182. doi: 10.1021/ja5009272

Figure 2.

Figure 2

Dph7 converts Δdph7 eEF2 to a substrate for amidation by Dph6. (A) Dph6 and Dph7 are both required for in vitro amidation of Δdph7 eEF2. The fluorescence labels indicate formation of diphthamide by the amidation reaction catalyzed by Dph6. (B) Stepwise in vitro reconstitution of diphthamide formation on Δdph7 eEF2. Lane 1: flag-tagged Δdph7 eEF-2 incubated without Dph7. Lane 2: flag-tagged Δdph7 eEF-2 incubated with Dph7 and then purified to remove Dph7. Both Δdph7 eEF-2 samples were then incubated with Dph6 and labeled with DT and Rh-NAD.