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. 2010 Jun 21;285(35):26779–26787. doi: 10.1074/jbc.M110.119693

FIGURE 3.

FIGURE 3.

Functional properties of eIF2D. A, lanes 1–7 show experiments analogous to those presented in Fig. 1A, but the purified recombinant factor was used instead of partially purified RSW fraction. Lanes 8–11 analyze the contribution of aminoacyl moiety of Met-tRNAiMet to its binding with 40 S ribosomal subunits. The HCV-UUU mRNA represents the HCV IRES where the initiation codon was mutated from AUG to UUU to test binding of cognate tRNAPhe in the presence of eIF2D (lanes 12–14). B, membrane binding assay. Recombinant eIF2D was tested for activity in [35S]Met-tRNAiMet binding with 40 S ribosomal subunits programmed with the AUG triplet. Bkg, background (no tRNA); control, no eIF2D added; eIF2A-1 and eIF2A-2 are two forms of eIF2A (see the text) purified to homogeneity.