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. 2015 Nov 18;3(2):e1117703. doi: 10.1080/21690731.2015.1117703

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© 2015 The Author(s). Published with license by Taylor & Francis

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 3. — The uS19 mutants promote increased affinity for TC and decreased affinity for the eEF2 translocase binding. (A) P-site tRNA binding. Salt-washed ribosomes were incubated for 10 minutes at 30ºC with two-fold dilutions of [¹⁴C]-ac-aa-tRNA and polyuridylic acid (Poly U). (B) A-site tRNA binding. Salt-washed ribosomes were incubated for 10 minutes at 30ºC with two-fold dilutions of pre-incubated ternary complex ([¹⁴C]-aa-tRNA•eEIF1A•GTP) and poly U. (C) eEF2 binding. Salt-washed ribosomes were incubated for 30 minutes at 30ºC with two-fold dilutions of purified eEF2, diphtheria toxin, and [¹⁴C]-NAD. In all 3 cases, binding was determined using liquid scintillation counting. (D) K_D values calculated using ligand depletion model in GraphPad Prism. *P < 0.05, **P < 0.01.