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. Author manuscript; available in PMC: 2010 Oct 30.
Published in final edited form as: J Mol Biol. 2009 Aug 14;393(3):608–618. doi: 10.1016/j.jmb.2009.08.018

Figure 4.

Figure 4

Supervillin L38K has significant and specific affinity for F-actin. (a) F-actin binding curves for villin headpiece (VHP, Kd = 6 μM), supervillin L38K (L38K, Kd = 88 μM), and wild-type supervillin headpiece (SVHP, Kd > 20,000 μM). (b) Results from F-actin competition assay performed between supervillin L38K and VHP. F-actin was incubated with a constant concentration of supervillin L38K (5 μM, diamonds; 25 μM, boxes; 100 μM, triangles) while the concentration of villin headpiece was increased from 0 to 200 μM. The amount of SVHP L38K bound to F-actin decreases with increasing concentrations of villin headpiece. The theoretical curves for pure competitive antagonism, calculated from equation 2 (See Materials and Methods) using the refined values calculated in (a), are drawn as solid lines. Inset: Representative data from F-actin competition assays performed between 100 μM SVHP L38K and increasing concentrations of VHP (5, 10, 25, 50, and 100 μM).