Refolding kinetics of cytochrome c studied with microsecond timescale continuous-flow UV-vis spectroscopy and rapid freeze-quench EPR

The refolding kinetics of acid denatured cytochrome c using the pH jump technique from pH 2 to pH 4.7 was studied using an ultrafast continuous flow mixing device that allowed time resolved measurements to the microsecond time scale. Three intermediate states were identified and characterized using UV-vis and EPR spectroscopy.137