Substrate water exchange in photosystem II core complexes of the extremophilic red alga Cyanidioschyzon merolae

Abstract

The binding affinity of the two substrate-water molecules to the water-oxidizing Mn₄CaO₅ catalyst in photosystem II core complexes of the extremophilic red alga Cyanidioschyzon merolae was studied in the S₂ and S₃ states by the exchange of bound ¹⁶O-substrate against ¹⁸O-labeled water. The rate of this exchange was detected via the membrane-inlet mass spectrometric analysis of flash-induced oxygen evolution. For both redox states a fast and slow phase of water-exchange was resolved at the mixed labeled m/z 34 mass peak: kf=52 ± 8s⁻¹ and ks=1.9 ± 0.3s⁻¹ in the S₂ state, and kf=42 ± 2s⁻¹ and kslow=1.2 ± 0.3s⁻¹ in S₃, respectively. Overall these exchange rates are similar to those observed previously with preparations of other organisms. The most remarkable finding is a significantly slower exchange at the fast substrate-water site in the S₂ state, which confirms beyond doubt that both substrate-water molecules are already bound in the S2 state. This leads to a very small change of the affinity for both the fast and the slowly exchanging substrates during the S₂→S₃ transition. Implications for recent models for water-oxidation are briefly discussed.

Keywords: Cyanidioschyzon merolae; Membrane-inlet mass spectrometry; Oxygen evolution; Photosystem II; Substrate–water exchange; Water oxidation.