Figure 3: NMR structure of the TTD bound to the Spacer.
From: Hemi-methylated DNA opens a closed conformation of UHRF1 to facilitate its histone recognition
(a) Ribbon representation of TTD–Spacer structure. N- and C-termini of the Spacer are indicated. The TTD is coloured in green, and the Spacer is coloured in yellow. The colour scheme is used the all the structure figures. (b) Superimposition of TTD–Spacer and TTD–PHD–H3K9me3 (4GY5.PDB) structures shown in ribbon representations. The TTD is coloured in green and the Spacer in yellow in TTD–Spacer structure. TTD–PHD–H3K9me3 complex is coloured in grey, and the PHD and H3K9me3 are omitted for simplicity. Residues for the interactions are shown in stick representation. (c) Electrostatic potential surface representation of the TTD with the Spacer shown in ribbon representation. The critical residues on the Spacer for the interaction are shown in stick representation. (d) Close-up view of TTD–Spacer interaction. Critical residues for the interaction are shown in stick representation. Hydrogen bonds are indicated as dashed lines. (e–g) Superimposed ITC enthalpy plots for the interaction between the Spacer and the TTD (or TTD–PHD) with the estimated binding affinity (KD) indicated. Wild-type and mutant proteins for the measurements are indicated. (h) GST pull-down assays for the intramolecular interactions. The wild-type or indicated truncations of UHRF1 were incubated with GST-tagged TTD, Linker or Spacer. The mixtures were immobilized on glutathione resin. The bound proteins were analysed by SDS–PAGE and Coomassie blue staining. ND, not determined; Sy., syringe.
