Figure 4.

Conformational Dynamics at Site II Depend on Binding Partners and Ligands

(A) Conformational equilibrium (73°C, 10 min) of the different ABCE1 site II states after incubation with saturating AMP-PNP concentrations (2 mM) and two representative 30S concentrations, as indicated.

(B) As in (A) with the full range of 30S concentrations. At each concentration, the percentage of every state was determined and plotted.

(C) Influence of different ligands such as nucleotides and ribosomal subunits on the conformational dynamics at site II. Measurements were performed under saturation conditions of nucleotides (2 mM) and 30S ribosomal subunits (1 μM) after 10 min of incubation at 73°C.

(D) Burst length distribution of the different conditions (as in C) were analyzed as previously described (Figures 2B and 2D).

(E) Time course experiment after incubation with saturating AMP-PNP (2 mM) and 30S (1 μM) concentrations at the representative points in time, as indicated (top 3 panels). Bottom panel: same experiment after pre-incubation with AMP-PNP (73°C, 2 mM, 8 min).

(F) As in (E), with the full range of points in time. At each point in time, the percentage of every state was determined and plotted.

(G) Cartoon summarizing the findings of (E) and (F). Data represent mean ± SD from 3–5 independent experiments.