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. 2016 May 16;36(11):1720–1732. doi: 10.1128/MCB.00097-16

FIG 5.

FIG 5

Experimental validation of the Ube2R1/2-acceptor ubiquitin complex confirming that Ile 44 on ubiquitin and Asp 91 on Ube2R2 do not help stabilize the interaction. (a) Time courses for diubiquitin synthesis comparing product formation for WT components with reaction mixtures containing WT Ube2R2 and I44R acceptor ubiquitin. (b) Same as in panel a, except D91K Ube2R2 activity has been assessed in the presence of WT or R54D acceptor ubiquitin. (c) Same as in panel a, except D58R acceptor ubiquitin activity has been assessed in the presence of either WT or D143K Ube2R2. (d) Same as in panel a, except R54A acceptor ubiquitin activity has been assessed in the presence of WT Ube2R2 or D143A Ube2R2 activity has been assessed in the presence of WT ubiquitin. Each experiment was performed in duplicate, and representative data are shown.