When iron(III) cytochrome c aqueous solutions containing NADH are irradiated with polychromatic light (wavelength greater than 280 nm), iron(II) cytochrome c and NAD+ in the stoichiometric ratio 2/1 are observed to be the principal reaction products, independently of the presence of oxygen; in addition, a minor process due to direct photodegradation of the nucleotide is observed. The selection of monochromatic 290 nm irradiation light (at which NADH has an absorbance minimum) and an adequate reactant concentration allowed parallel reactions to be minimized and new information to be obtained on the mechanism of the photoredox process. The experimental results are consistent with a reaction mechanism whereby NADH donates one electron to a "reactive intermediate" of the hemoprotein formed from the light-induced methionine-to-iron charge transfer excited state. In this process an NAD. radical is formed which, in deaerated solution, immediately reduces another molecule of the hemoprotein, and is itself oxidized to NAD+. In aerated solution, the NAD. radical rapidly reacts with oxygen to give NAD+ and superoxide O2- anion radical which, in turn, reduces the second iron(III) cytochrome c molecule.
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