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. 2005 Nov 17;24(24):4316–4323. doi: 10.1038/sj.emboj.7600884

Figure 3.

Figure 3

Effect of L7/12 mutations on single-round GTP hydrolysis and translocation catalyzed by EF-G. (A) Time courses of GTP hydrolysis. EF-G and [γ-32P]GTP were rapidly mixed with ribosomes (Materials and methods). Unlabeled GTP (1 mM) was added together with the ribosomes in order to limit [γ-32P]GTP hydrolysis to a single round (Rodnina et al, 1997). Symbols as in Figure 2. (B) Rate constants of GTP hydrolysis. Time courses shown in panel A were evaluated by exponential fitting to yield rate constants as indicated. (C) Time courses of translocation as monitored by the fluorescence of fMetPhe-tRNAPhe(Prf16/17). Overlaid stopped-flow traces are shown for ribosomes reconstituted with wt L7/12, V66A, V66D, I69A, K70A, R73M, and R73K.