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. 2016 Jun 16;17(8):1221–1235. doi: 10.15252/embr.201642641

Figure EV3. HHARI RING1 disruption of (E3‐independent) closed E2~Ub conformations.

Figure EV3

Similar to UbcH7, Ubc13~Ub populates closed states in the absence of an E3 35. Shown here are three 1H‐15N‐HSQC‐TROSY spectra of Ub overlaid: free 15N‐Ub (blue), Ubc13‐O‐15N‐Ub (black), and Ubc13‐O‐15N‐Ub in the presence of 3 mol equiv. HHARI RING1 (red). Due to the weak affinity binding of Ubc13 to HHARI RING1, the complex is far from saturation and the spectrum exhibits fast‐to‐intermediate exchange behavior. Therefore, the observed CSPs for the effects of RING1 binding to Ubc13‐O‐15N‐Ub are quite small (black versus red). Black arrows highlight several Ub resonances that shift along the same trajectory (but opposite direction) upon binding HHARI RING1 as experienced when Ub is conjugated to Ubc13, consistent with Ubc13~Ub populating more open‐like states when in complex with HHARI RING1.