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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Nat Chem Biol. 2014 Dec 1;11(1):83–89. doi: 10.1038/nchembio.1700

Figure 6.

Figure 6

The Ube2w C-terminus facilitates α-amino group reactivity. (a) Selected {1H – 15N} resonances of residues near the Ube2w active site are compared in the spectra of full-length WT (black), W144E-Ube2w, (purple), and Ube2w-131Δ (red). Resonances move along similar trajectories as a result of the W144 mutation or C-terminal ablation, indicating similar chemical environments for the affected residues. (b) Mutation or ablation of the C-terminus affects the intrinsic aminolysis activity of Ube2w. In a 1hr reaction WT Ube2w~Ub shows robust transfer activity towards peptide (NH2-A-G-G-S-Y-COO-; 30 mM) as seen by increased amounts of free Ube2w and free Ub. Ube2w-W144E~Ub and the Ube2w-131Δ~Ub mutants show almost no Ub transfer activity to this minimal substrate over the same time period (Supplementary Fig. 2l).