Figure 9. Phosphomimetic mutation S80E displaces the Box from the RING and allows E2 enzyme binding.

(A) Comparison of chemical shift perturbations upon E2 enzyme Ube2N titration into T21 RB (grey) or T21 RB S80E (black). E2 interacting regions are highlighted in green and secondary structure is indicated by rectangles (helices) and arrows (β-strands). (B) Comparison of heteronuclear NOEs for T21 RB (grey) or T21 RB S80E (black) in absence of E2. The data is consistent with increased dynamics in the linker region between the RING and the Box upon introduction of S80E. (C) Relative solvent exposure in S80E vs. WT, as detected by solvent PREs (shown as intensity ratio WT/S80E mapped onto the RB structure; primary data in Supplementary file 2). White areas indicate regions more solvent-exposed in WT while red areas indicate parts of the surface more solvent-exposed in S80E.

Figure 9—figure supplement 1. Main chain temperature factors for T21 RING-Box structure.