Proton and Water Transfer Pathways in the S₂ → S₃ Transition of the Water-Oxidizing Complex in Photosystem II: Time-Resolved Infrared Analysis of the Effects of D1-N298A Mutation and NO₃^– Substitution

Abstract

Photosynthetic water oxidation is performed through a light-driven cycle of five intermediates (S₀-S₄ states) in photosystem II (PSII). The S₂ → S₃ transition, which involves concerted water and proton transfer, is a key process for understanding the water oxidation mechanism. Here, to identify the water and proton transfer pathways during the S₂ → S₃ transition, we examined the effects of D1-N298A mutation and NO₃^- substitution for Cl^-, which perturbed the O1 and Cl channels, respectively, on the S₂ → S₃ kinetics using time-resolved infrared spectroscopy. The S₂ → S₃ transition was retarded both upon NO₃^- substitution and upon D1-N298A mutation, whereas it was unaffected by further NO₃^- substitution in N298A PSII. The H/D kinetic isotope effect in N298A PSII was relatively small, revealing that water transfer is a rate-limiting step in this mutant. From these results, it was suggested that during the S₂ → S₃ transition, water delivery and proton release occur through the O1 and Cl channels, respectively.