Fig. 3.

Biophysical characterization of the designed and WT acylphosphatase (AcPh) and Cdcd42. (A) Comparison of the experimental partial molar heat capacity profiles of AcPh-wt (white circles) and AcPh-des (red squares). Solid lines are the results of the fit according to a 2-state unfolding model. The results of the fit are given in Table 1. (B) Comparison of the experimental temperature-induced unfolding profiles of AcPh-wt (black thin line) and AcPh-des (red thin line) as monitored by the changes in ellipticity at 222 nm. Thick solid lines are the results of the fit according to a 2-state unfolding model. The results of the fit are given in Table 1. (C) Comparison of the experimental temperature induced unfolding profiles of Cdc42-wt (thin black line), Cdc42-des1 (thin blue line), and Cdc42-des2 (thin red line) as monitored by the changes in ellipticity at 222 nm. (D) Dependence of specific activity of Cdc42 variants (Cdc42-wt, black circles; Cdc42-des1, blue squares; Cdc42-des2, red triangles) on temperature. Lines are drawn to guide the eye. (E) Resistance of Cdc42 variants to aggregation after exposure to high temperature. The fraction of soluble monomer remained in solution after exposure to elevated temperatures was measured by using size-exclusion chromatography as described in Materials and Methods. Shown are Cdc42-wt (black circles), Cdc42-des1 (blue squares), and Cdc42-des2 (red triangles), and lines are drawn to guide the eye.