Melting temperatures and thermodynamic parameters for urea-induced unfolding equilibrium of Pim-1 wild type and mutants measured by far-UV CD spectroscopy.

The temperature-induced changes were followed by monitoring the ellipticity at 209 nm. The Tm values were calculated by taking the first derivative of the ellipticity at 209 nm with respect to temperature. Tm1 and Tm2 refer to the first and second transition observed for wild type and Y53H, respectively. For urea-induced unfolding equilibrium the data were obtained at 10°C in 20 mM Tris/HCl, pH 7.5, containing 0.2 M NaCl and 200 µM DTT by measuring circular dichroism ellipticity at 222 nm [Θ222]. ΔGH2O and m values were obtained from Eqn. 3; [Urea]0.5 was calculated from Eqn. 4. Data are reported as the mean ± SE of the fit.