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. Author manuscript; available in PMC: 2008 Oct 26.
Published in final edited form as: J Mol Biol. 2007 Aug 2;373(3):562–572. doi: 10.1016/j.jmb.2007.07.032

Figure 7. Direct comparison of rapid kinetic measures accompanying 70SIC formation.

Figure 7

All concentrations are final after mixing. In each experiment, 50S concentration was 3 μM. IF2c fluorescence corrected for light scattering (red traces); Concentrations of 30SIC components were: 30S, 0.3 μM; IF1, IF3 and fMet-tRNAfMet, 0.45 μM; IF2C, 0.15 μM; AUG022mRNA, 0.9 μM; GTP, 100 μM. Light scattering (blue traces); concentrations were the same as for IF2C fluorescence, with IF2 replacing IF2C. GTPase (black traces and solid circles); concentrations were the same as for light scattering with the following differences: 30S, 0.45 μM; IF2, 0.3 μM; GTP, 36 μM.; Pi release (green traces); concentrations were the same as for light scattering except for IF2 (0.45 μM). fMet-tRNAfMet(prf 20) fluorescence (orange traces); concentrations are the same as for Pi release, but with fMet-tRNAfMet (prf20) 0.18μM, replacing fMet-tRNAfMet. (A) Early time period; (B) Extended time period. For convenience of presentation, the rapid initial phase of fMet-tRNAfMet (prf20) fluorescence change on mixing with 50S subunits is not shown. The plateau stoichiometries of Pi formation and release were both 0.8 ± 0.1/IF2. Solid lines through experimental traces are the results of global fitting to Scheme 2.