Flash-induced redox changes in oxygen-evolving spinach Photosystem II core particles

Abstract

Flash-induced redox reactions in spinach PS II core particles were investigated with absorbance difference spectroscopy in the UV-region and EPR spectroscopy. In the absence of artificial electron acceptors, electron transport was limited to a single turnover. Addition of the electron acceptors DCBQ and ferricyanide restored the characteristic period-four oscillation in the UV absorbance associated with the S-state cycle, but not the period-two oscillation indicative of the alternating appearance and disappearance of a semiquinone at the Q_B-site. In contrast to PS II membranes, all active centers were in state S₁ after dark adaptation. The absorbance increase associated with the S-state transitions on the first two flashes, attributed to the Z⁺S₁→ZS₂ and Z⁺S₂→ZS₃ transitions, respectively, had half-times of 95 and 380 μs, similar to those reported for PS II membrane fragments. The decrease due to the Z⁺S₃→ZS₀ transition on the third flash had a half-time of 4.5 ms, as in salt-washed PS II membrane fragments. On the fourth flash a small, unresolved, increase of less than 3 μs was observed, which might be due to the Z⁺S₀→ZS₁ transition. The deactivation of the higher S-states was unusually fast and occurred within a few seconds and so was the oxidation of S₀ to S₁ in the dark, which had a half-time of 2–3 min. The same lifetime was found for tyrosine D⁺, which appeared to be formed within milliseconds after the first flash in about 10% inactive centers and after the third and later flashes by active centers in Z⁺S₃.