Figure 5.

Temperature dependence of the refolding of intact CI-2, the association of CI2(1–40) and CI2(41–64) fragments, and renaturation of pressure denatured state. (a) The major rate of folding of acid-unfolded CI-2 (10 μM) was measured at different temperatures, and the data represented in the figure were fitted to a nonlinear Eyring equation (cf. ref. 23). (b) CI2(1–40) (1 μM) was mixed with 10 μM of CI2(41–64), the fluorescence change was monitored at different temperatures, and the rate of the major phase represented and fitted as in a. (c) The complex (10 μM) was denatured by pressure at the indicated temperatures until the center of spectral mass was equilibrated at the corresponding “denatured” value. The pressure was released and the fluorescence was monitored. Data for each temperature were analyzed as two exponentials as in Fig. 1 and the observed rates of the major phase plotted. The residuals of the single-exponential fitting are shown at the bottom of each plot.