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. 2021 Mar 30;10:e66151. doi: 10.7554/eLife.66151

Figure 4. Stabilizing the ‘threonine belt’ in the R8 domain of talin inhibits talin-vinculin interactions under tension-free conditions.

(a) Cartoon representation of talin R7R8 (pdb id 2X0C) showing the ‘threonine belt’, comprised of residues T1502, T1542, and T1562, labeled and shown as sticks (cyan), the VBS helix is colored red. (a.i) side on view (N.B. helix 31 transparent), (a.ii) top down view. (b) Denaturation profiles for WT R7R8wt (red) and R7R8vvv (black) measured by monitoring the change in circular dichroism at 208 nm with increasing temperature. R7R8wt has a melting temperature of 55°C, whereas R7R8vvv unfolds in two steps, one (R7) with a melting temperature of 56°C and R8 unfolding at 82°C. (c) Chromatograms showing binding of talin R7R8 to the vinculin head (Vd1). R7R8wt (red) and R7R8vvv (black) binding to Vd1. Complex peaks and unbound peaks are indicated.

Figure 4.

Figure 4—figure supplement 1. Fluorescence polarization assay showing the binding affinities for R8 ligand peptides from (top) RIAM TBS1 and (bottom) DLC1 with WT and R7R8vvv.

Figure 4—figure supplement 1.