FIGURE 5.

Physical isolation of ternary complex. Ternary complex formed in the presence of saturating concentrations of EF-Tu (circles) or EF-Tu·EF-Ts (diamonds), Cy3-labeled Phe-tRNA^Phe, and GTP (see “Experimental Procedures”) was fractionated over a Superdex 75 gel filtration column in the presence (black) or absence (red) of GTP (10 μm) in the running buffer. Absorbance was recorded at 550 and 260 nm to specifically track the elution times of “unbound” and ternary complex-bound Cy3-labeled Phe-tRNA^Phe as indicated. This was repeated with GDPNP in the reaction and running buffer (blue triangles).