Figure 1
From: Angiomotin binding-induced activation of Merlin/NF2 in the Hippo pathway
Mammalian Merlin adopts a semi-open conformation. (A) Schematic diagrams showing the domain organizations of Merlin and EBP50. The Merlin-FERM/EBP50-CT interaction is indicated by a two-way arrow. The S518 site is indicated by a red asterisk and used throughout the rest of the figures. The color coding scheme of Merlin is used throughout the rest of the paper unless specified otherwise. (B-D) ITC-based measurements of the bindings between EBP50-CT and Merlin-FERM (B), WT-Merlin and various mutant forms of full-length Merlin (C, D). (E) Sedimentation velocity analysis showing the hydrodynamic properties of WT Merlin and A585W-Merlin. The fitted molecular weight of A585W-Merlin is indicated. The green dashed line scales the y-axis of the WT-Merlin profile by 10 folds to show that the broader peak at S value ∼2.5 when compared to that of A585W-Merlin. The loading concentrations of the two proteins for the experiment are the same. (F, G) Differential scanning fluorimetry-based thermal denaturation assay showing the temperature-dependent denaturation profiles of WT-Merlin, S518D-Merlin, A585W-Merlin, and S518D/A585W-Merlin.
