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 (U1(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.