Interrupted refolding of CMPK wt.

(a) Mixing scheme of the interrupted refolding reaction. After refolding in 1.2 M urea for incubation time t1, the protein is diluted back into 6.0 M urea. The subsequent unfolding is recorded as function of unfolding time t2 (b). For short incubation times (<100 s), a transient fast unfolding phase λU1(IR) can be observed. For long incubation times, the slow λU3(IR) unfolding phase also described in the single jump experiments appears. A secondary plot of the amplitudes AU1(IR) (•) and AU3(IR) (□) corresponding to the rate constants λU1(IR) and λU3(IR) is shown in (c). A global fit of this data yields secondary rate constants of ΛF1(IR) = 5.9 s−1 and ΛF3(IR) = 0.0046 s−1.