Identification of Regions Involved in Substrate Binding and Dimer Stabilization within the Central Domains of Yeast Hsp40 Sis1
Figure 3
Urea- and thermal-induced unfolding measurements followed by CD.
The recombinant proteins (0.5 mg/mL) were submitted to thermal-unfolding followed by CD at 222 nm. (A) Urea-induced unfolding experiments were followed by CD at 222 nm, using a 1 mm pathlength cell in buffer A, at 20°C after 90–120 min of equilibration. Data are shown as fraction of unfolded protein and represent the mean of three independent experiments. The three-state unfolding transition model (Eq. 1) was used to fit the fraction of unfolding data and the fitting is shown by the line (see text for details). The unfolding experiments (in triplicate) were measured from 20°C to 90°C with a scan rate of 1.0°C/min. Data is presented as [θ] (B) and in delta of signal (C), which was fitted using a sigmoidal function (full line) yielding the TmCD.