Oxidation-reduction potentials of bound iron-sulfur proteins of photosystem I.

Digitonin - fractionated photosystem - I subchloroplasts were titrated potentiometrically between -450 and -610 mV at pH 10. Examination of the titrated subchloroplasts by low-temperature (13 degrees K) electron paramagnetic resonance spectroscopy revealed resonances centered at values of 2.05, 1.94, 1.92, 1.89, and 1.86 on the g-factor scale. The peak heights depended on the potentials at which the chloroplasts were poised. The resonances of at least three iron-sulfur centers can be recognized: one with lines at g = 2.05 and 1.94; one with lines at g = 2.05, 1.92, and 1.89; and one for which only a line at g = 1.86 has been resolved. The midpoint potentials of the iron-sulfur species fall into two distinctly separate regions: the titration profile of the g = 1.94 signal, the first segment of the g = 2.05 plot, and the rise phase of the g = 1.86 signal had a value of -530 +/- 5 mV; the upper segment of the g = 2.05 plot, the decrease phase of the g = 1.86 signal, and the g = 1.89 profile had a midpoint potential estimated to be [unk] -580 mV. The oxidation-reduction reaction of each of the bound iron-sulfur species, as represented by the changes of the electron paramagnetic resonance spectra, was reversible and apparently involved a two-electron change.Titration at pH 9 could only be carried to -560 mV, and essentially only the first half of the titration behavior as found at pH 10 was seen. At any given potential more positive than -560 mV, the part of the iron-sulfur protein that was not reduced electrochemically could be reduced photochemically, but only to the maximum extent reduced electrochemically at -560 mV. Whereas, chloroplasts illuminated at room temperature and then frozen while still being illuminated developed a signal similar to that produced by electrochemical reduction at -610 mV, illumination at 77 degrees K did not bring about photoreduction beyond that accomplished electrochemically at about -560 mV.Dithionite alone in the dark and under anaerobic conditions brought about a partial reduction to the extent of the first electrochemical reduction step. Dithionite plus illumination at room temperature or dithionite plus methyl viologen in the dark produced the maximum signal. Electron paramagnetic resonance spectra due to either light or electrochemically reduced iron-sulfur proteins showed no detectable decay for at least 3 days when samples were stored in the dark at 77 degrees K.