Proton Translocation via Tautomerization of Asn298 During the S₂–S₃ State Transition in the Oxygen-Evolving Complex of Photosystem II

In biological water oxidation, a redox-active tyrosine residue (D1-Tyr161 or Y_Z) mediates electron transfer between the Mn₄CaO₅ cluster of the oxygen-evolving complex and the charge-separation site of photosystem II (PSII), driving the cluster through progressively higher oxidation states S_i (i = 0–4). In contrast to lower S-states (S₀, S₁), in higher S-states (S₂, S₃) of the Mn₄CaO₅ cluster, Y_Z cannot be oxidized at cryogenic temperatures due to the accumulation of positive charge in the S₁ → S₂ transition. However, oxidation of Y_Z by illumination of S₂ at 77–190 K followed by rapid freezing and charge recombination between Y_Z^• and the plastoquinone radical Q_A^•– allows trapping of an S₂ variant, the so-called S₂^trapped state (S₂^t), that is capable of forming Y_Z^• at cryogenic temperature. To identify the differences between the S₂ and S₂^t states, we used the S₂^tY_Z^• intermediate as a probe for the S₂^t state and followed the S₂^tY_Z^•/Q_A^•– recombination kinetics at 10 K using time-resolved electron paramagnetic resonance spectroscopy in H₂O and D₂O. The results show that while S₂^tY_Z^•/Q_A^•– recombination can be described as pure electron transfer occurring in the Marcus inverted region, the S₂^t → S₂ reversion depends on proton rearrangement and exhibits a strong kinetic isotope effect. This suggests that Y_Z oxidation in the S₂^t state is facilitated by favorable proton redistribution in the vicinity of Y_Z, most likely within the hydrogen-bonded Y_Z–His190–Asn298 triad. Computational models show that tautomerization of Asn298 to its imidic acid form enables proton translocation to an adjacent asparagine-rich cavity of water molecules that functions as a proton reservoir and can further participate in proton egress to the lumen.