Figure 4. Varied auto inhibition of DIAPH1-FFC mutants.
Pyrene-actin assembly assays were carried out with 2 μM actin (5% pyrene-labeled), ± 5 nM of FFC, either (A) WT; (B) ttaa ;(C) 1213x; (D) Δag; or (E) M1199D, with varied concentration of WT DIAPH1-NT. Concentrations of NT are indicated in (A), and the color scheme is the same in panels (B-E), with actin plus 200 nM NT and no FFC in gray. (F) Inhibition curves were calculated from the slopes of raw pyrene traces at 100 s and fit with a quadratic binding model that assumes 1:1 binding between dimers. A replicate of this experiment with completely different protein preparations is shown in Figure S2.