Interrupted unfolding of CMPK wt.

The mixing-scheme of the interrupted unfolding reaction is displayed in (a). After unfolding in 6.0 M urea with incubation time t1, the protein is diluted into 1.2 M urea. The subsequent refolding is recorded as function of refolding time t2 (b). For short incubation times (<100 s), a transient intermediate refolding phase λF2(IU) can be observed. For long incubation times, the fast and slow refolding phases λF1(RS) and λF3(RS) described in the single jump experiments sufficiently describe the observed transitions λF1(IU) and λF3(IU). A secondary plot of the amplitudes AF1(IU) (•), AF2(IU) (□) and AF3(IU) (×) corresponding to the rate constants λF1(IU), λF2(IU) and λF3(IU) is shown in (c). A global fit performed on these amplitudes yields rate constants of ΛU2(IU) = 0.14 s−1 and ΛU3(IU) = 0.012 s−1. The fits are depicted as solid (AF1(IU)), dashed (AF2(IU)) and dotted (AF3(IU)) lines. To account for the different scales in amplitude changes, the y-axis is splitted at 0.1 a.u.