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. 2016 Mar 9;291(19):10426–10436. doi: 10.1074/jbc.M115.697789

FIGURE 4.

FIGURE 4.

Effects of CCA addition and charging on the interaction of tRNA with cytochrome c. A, kinetics of A76 addition to etRNAVal (1 μm) catalyzed by human CCA enzyme (2 μm) at 37 °C in the presence or absence of cytochrome c (15 μm). B, ability of free etRNACys and etRNACys in complex with CCA-adding enzyme to bind cytochrome c. Analysis of fluorescence quenching as a function of cytochrome c concentration in the fluorescence-based assay. Error bars indicate S.D. C, model of tRNA in complex with CCA-adding enzyme of Archaeoglobus fulgidus (green, Protein Data Bank code 1sz1) and CysRS of E. coli (gray, Protein Data Bank code 1u0b), showing the capacity to accommodate three molecules of cytochrome c (brown, purple, and pink, Protein Data Bank code 3ZCF) on the outside corner of the L-structure. D, dissociation constant (Kd) of cytochrome c with uncharged and charged Cy3-etRNACys. Error bars indicate S.D. E, ability of free etRNACys or tRNACys in complex with CysRS to bind cytochrome c. Analysis of fluorescence quenching as a function of cytochrome c concentration in the fluorescence-based assay. Error bars indicate S.D.