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. 2006 May 25;25(11):2539–2550. doi: 10.1038/sj.emboj.7601140

Figure 2.

Figure 2

Effect of initiation factors on fMet-tRNAfMet binding to 30S ribosomal subunits. (A) [3H]fMet-tRNAfMet association to mRNA-programmed 30S subunits was measured by NC filtration technique in the absence of initiation factors or with only IF1 present. Insert: dependence of fMet-tRNAfMet association rate on the concentration of fMet-tRNAfMet in the absence of initiation factors. (B) [3H]fMet-tRNAfMet dissociation from 30S preinitiation complexes assembled with different combinations of IF1 and IF2 was measured by NC filtration in the presence of a 10-fold excess of nonlabeled chasing fMet-tRNAfMet. (C) The same as in (B) except that IF3 was also present in the 30S preinitiation complex. (D) Dependence of the 30S·mRNA·IF3·fMet-tRNAfMet complex formation on the concentration of fMet-tRNAfMet in the presence or absence of IF1 measured by NC filtration. (E) Kinetics of fMet-tRNAfMet binding to 30S·mRNA·IF2 or to 30S·mRNA·IF1·IF2 complexes measured by light scattering. (F) The same as in (E) but in the presence of IF3.