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. 2019 Nov 7;9:16195. doi: 10.1038/s41598-019-52618-3

Figure 3.

Figure 3

TbUBA1a and TbUBA1b are resistant to TAK-243 inhibition. (A) E1-E2 transthioesterification reactions with the indicated UBA1s in the absence (0) or presence of increasing concentrations of TAK-243. Note the difference in concentrations of TAK-243 used for hUBA1 on the one hand, and TbUBA1a and TbUBA1b on the other hand. Reactions were stopped after 30 min. E1 activity was determined by the transfer of ubiquitin to the E2 (UbcH5a). Indicated are free E2 (E2) and the E2-ubiquitin conjugate (E2~Ub). Images are derived from the same Coomassie stained gel. See Supplementary Information for an image of the full length gel. (B) Quantification of the experiment shown in (A). E2~Ub was calculated as a percentage of the total amount of E2 [E2~Ub/(E2 + E2~Ub)]. This was set at 100% for the reaction in the absence of TAK-243 (0) for each protein separately. (C) E1-E2 transthioesterification reactions as in (A) performed in the absence (0) or presence of increasing concentrations of PYR-41. Images are derived from the same Coomassie stained gel. See Supplementary Information for an image of the full length gel.