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. 2014 Jul 15;5:4395. doi: 10.1038/ncomms5395

Figure 4. Time-resolved absorbance changes upon laser excitation at pH 5.8 in the absence of nitrite.

Figure 4

Absorbance changes at 595 nm observed in the laser flash-photolysis instrument during reduction of wild-type (WT) AxNiR (black) and F306C (red) with NADH (λexc=355 nm). Experiments were carried out in the absence of nitrite in MMH buffer, pH 5.8, containing 30 μM WT AxNiR or F306C, NiR, 200 μM NADH and 50 mM mediator at 4 °C. Traces shown are the average of 3–6 individual traces and were fitted by using a double-exponential equation. Observed rates of inter-Cu ET for WT AxNiR and F306C are 200 and 160 s−1, respectively. The lower absorbance at equilibrium observed for F306C is consistent with an ~60 mV decrease in the Em of the T2Cu centre.