On the simulation and interpretation of substrate-water exchange experiments in photosynthetic water oxidation.

Abstract

Water oxidation by photosystem II (PSII) sustains most life on Earth, but the molecular mechanism of this unique process remains controversial. The ongoing identification of the binding sites and modes of the two water-derived substrate oxygens ('substrate waters') in the various intermediates (S_i states, i = 0, 1, 2, 3, 4) that the water-splitting tetra-manganese calcium penta-oxygen (Mn₄CaO₅) cluster attains during the reaction cycle provides central information towards resolving the unique chemistry of biological water oxidation. Mass spectrometric measurements of single- and double-labeled dioxygen species after various incubation times of PSII with H₂¹⁸O provide insight into the substrate binding modes and sites via determination of exchange rates. Such experiments have revealed that the two substrate waters exchange with different rates that vary independently with the S_i state and are hence referred to as the fast (W_f) and the slow (W_S) substrate waters. New insight for the molecular interpretation of these rates arises from our recent finding that in the S₂ state, under special experimental conditions, two different rates of W_S exchange are observed that appear to correlate with the high spin and low spin conformations of the Mn₄CaO₅ cluster. Here, we reexamine and unite various proposed methods for extracting and assigning rate constants from this recent data set. The analysis results in a molecular model for substrate-water binding and exchange that reconciles the expected non-exchangeability of the central oxo bridge O5 when located between two Mn(IV) ions with the experimental and theoretical assignment of O5 as W_S in all S states. The analysis also excludes other published proposals for explaining the water exchange kinetics.