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. 2015 Aug 7;34(20):2506–2521. doi: 10.15252/embj.201592337

Figure 3.

Figure 3

The RING0/RING1 interface is a hinge that remodels upon parkin phosphorylation
  1. Overlay of UblR0RBR (coloured according to Fig1C) and R0RBR (grey) structures. Absence of the Ubl domain causes a hinge opening at the RING0/RING1 interface.
  2. Close-up of the RING0/RING1 interface in UblR0RBR showing key residues that are involved in ionic/hydrogen bond interactions.
  3. Overlay of the RING0/RING1 interface from UblR0RBR (coloured) and R0RBR (grey). Green dashes show distances in Å between residues in UblR0RBR, and red dashes show distances between residues in R0RBR.
  4. Structure of S65DUblR0RBR (pink) overlaid with UblR0RBR parkin (coloured according to Fig1C). The position of D65 or S65 is shown in inset.
  5. Comparison of the hinge in UblR0RBR (left), S65DUblR0RBR (middle) and R0RBR (right).
  6. Surface of UblR0RBR (left) and S65DUblR0RBR (right) showing continuous basic patch formed by hinge opening.
  7. Ubiquitination assays of mutations in the RING0/RING1 interface. Mutations in residues that pin H302 in the flipped-in position are activating, mutation of the basic patch prevents parkin activity, even in the presence of increasing concentrations of phosphoubiquitin (+). This is the case for both chain formation (top) and Miro1 ubiquitination (bottom). A Coomassie-stained loading control is shown in between. A non-specific, ATP-independent band is indicated (*).