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. 2018 Jan 4;46(3):1395–1411. doi: 10.1093/nar/gkx1300

Figure 5.

Figure 5.

Binding of tRNAThr by the TsaB homodimer, TsaD homodimer, TsaE and pre-isolated complexes. Proteins were incubated with 100 nM of 32P-labeled tRNAThr at room temperature and the mixtures separated by native PAGE (see ‘Materials and Methods’ section). (A) Autoradiograms showing unbound tRNAThr (at the bottom of each gel) and the nucleoprotein complexes (at the top of each gel). (B) Plot of fraction of tRNA bound quantified by densitometry as the ratio of upper-band intensity to total intensity in each lane. (C) TsaE-dependent release of tRNA from the TsaB2D2 complex. TsaB2D2 complex (5 μM) pre-incubated with 32P-labeled tRNAThr (100 nM) at room temperature was titrated with increasing concentrations of TsaE, and analyzed by native PAGE. Top panel: autoradiogram of the native gel showing free and bound tRNA at the bottom and top of the gel, respectively. Bottom panel: coomassie staining of the same gel. TsaE concentrations are indicated at the top, and lane numbers are indicated at the bottom. (D) Densitometric quantification of the radioactive bands in panel A, n = 2.