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. Author manuscript; available in PMC: 2016 May 15.
Published in final edited form as: Biochem J. 2015 May 15;468(1):25–31. doi: 10.1042/BJ20141410

Figure 4. Transient kinetics of POR reduction by one molar equivalent of NADPH.

Figure 4

(A) Electron transfer scheme. (B) and (C) Absorbance decay at 450 nm, reflecting flavin reduction, which includes FAD reduction and also, to a lesser extent, electron transfer from FADH2 to FMN catalysed by WT and A287P POR. (D) and (E) Absorbance increase at 590 nm, reflecting the formation of the blue disemiquinone (FADH•/FMNH•) catalysed by WT and A287P POR. Black lines show best fits to the normalized data. Equal molar amounts of POR and NADPH solutions were rapidly mixed and changes in absorbance at 450 nm and 590 nm were followed using a stopped-flow spectrometer. Final concentrations after mixing were 15.98 µM for WT and 17.7 µM for A287P.