Insights into substrate binding to the oxygen-evolving complex of photosystem II from ammonia inhibition studies

© 2014 American Chemical Society. Water oxidation in Photosystem II occurs at the oxygen-evolving complex (OEC), which cycles through distinct intermediates, S0-S4. The inhibitor ammonia selectively binds to the S2 state at an unresolved site that is not competitive with substrate water. By monitoring the yields of flash-induced oxygen production, we show that ammonia decreases the net efficiency of OEC turnover and slows the decay kinetics of S2 to S1. The temperature dependence of biphasic S2 decay kinetics provides activation energies that do not vary in control and ammonia conditions. We interpret our data in the broader context of previous studies by introducing a kinetic model for both the formation and decay of ammonia-bound S2. The model predicts ammonia binds to S2 rapidly (t1/2 = 1 ms) with a large equilibrium constant. This finding implies that ammonia decreases the reduction potential of S2 by at least 2.7 kcal mol-1 (\u3e120 mV), which is not consistent with ammonia substitution of a terminal water ligand of Mn(IV). Instead, these data support the proposal that ammonia binds as a bridging ligand between two Mn atoms. Implications for the mechanism of O-O bond formation are discussed

Photosynthetic water oxidation: Binding and activation of substrate waters for O-O bond formation

© 2015 The Royal Society of Chemistry. Photosynthetic water oxidation occurs at the oxygen-evolving complex (OEC) of Photosystem II (PSII). The OEC, which contains a Mn4CaO5 inorganic cluster ligated by oxides, waters and amino-acid residues, cycles through five redox intermediates known as Si states (i = 0-4). The electronic and structural properties of the transient S4 intermediate that forms the O-O bond are not well understood. In order to gain insight into how water is activated for O-O bond formation in the S4 intermediate, we have performed a detailed analysis of S-state dependent substrate water binding kinetics taking into consideration data from Mn coordination complexes. This analysis supports a model in which the substrate waters are both bound as terminal ligands and react via a water-nucleophile attack mechanism

Desiccation tolerant lichens facilitate in vivo H/D isotope effect measurements in oxygenic photosynthesis

© 2018 Elsevier B.V. We have used the desiccation-tolerant lichen Flavoparmelia caperata, containing the green algal photobiont Trebouxia gelatinosa, to examine H/D isotope effects in Photosystem II in vivo. Artifact-free H/D isotope effects on both PSII primary charge separation and water oxidation yields were determined as a function of flash rate from chlorophyll-a variable fluorescence yields. Intact lichens could be reversibly dehydrated/re-hydrated with H2O/D2O repeatedly without loss of O2 evolution, unlike all isolated PSII preparations. Above a threshold flash rate, PSII charge separation decreases sharply in both D2O and H2O, reflecting loss of excitation migration and capture by PSII. Changes in H/D coordinates further slow charge separation in D2O (−23% at 120 Hz), attributed to reoxidation of the primary acceptor QA−. At intermediate flash rates (5–50 Hz) D2O decreases water oxidation efficiency (O2 evolution) by −2–5%. No significant isotopic difference is observed at slow flash rates (\u3c5 \u3eHz) where charge recombination dominates. Slower D2O diffusion, changes in hydrogen bonding networks, and shifts in the pKa\u27s of ionizable residues may all contribute to these systematic variations of H/D isotope effects. Lichens’ reversible desiccation tolerance allows highly reproducible H/D exchange kinetics in PSII reactions to be studied in vivo for the first time

A multi-iron system capable of rapid N\u3csub\u3e2\u3c/sub\u3e formation and N \u3csub\u3e2\u3c/sub\u3e cleavage

The six-electron oxidation of two nitrides to N2 is a key step of ammonia synthesis and decomposition reactions on surfaces. In molecular complexes, nitride coupling has been observed with terminal nitrides, but not with bridging nitride complexes that more closely resemble catalytically important surface species. Further, nitride coupling has not been reported in systems where the nitrides are derived from N2. Here, we show that a molecular diiron(II) diiron(III) bis(nitride) complex reacts with Lewis bases, leading to the rapid six-electron oxidation of two bridging nitrides to form N2. Surprisingly, these mild reagents generate high yields of iron(I) products from the iron(II/III) starting material. This is the first molecular system that both breaks and forms the triple bond of N2 at room temperature. These results highlight the ability of multi-iron species to decrease the energy barriers associated with the activation of strong bonds. © 2014 American Chemical Society

Thermodynamically accurate modeling of the catalytic cycle of photosynthetic oxygen evolution: A mathematical solution to asymmetric Markov chains

AbstractForty-three years ago, Kok and coworkers introduced a phenomenological model describing period-four oscillations in O2 flash yields during photosynthetic water oxidation (WOC), which had been first reported by Joliot and coworkers. The original two-parameter Kok model was subsequently extended in its level of complexity to better simulate diverse data sets, including intact cells and isolated PSII-WOCs, but at the expense of introducing physically unrealistic assumptions necessary to enable numerical solutions. To date, analytical solutions have been found only for symmetric Kok models (inefficiencies are equally probable for all intermediates, called “S-states”). However, it is widely accepted that S-state reaction steps are not identical and some are not reversible (by thermodynamic restraints) thereby causing asymmetric cycles. We have developed a mathematically more rigorous foundation that eliminates unphysical assumptions known to be in conflict with experiments and adopts a new experimental constraint on solutions. This new algorithm termed STEAMM for S-state Transition Eigenvalues of Asymmetric Markov Models enables solutions to models having fewer adjustable parameters and uses automated fitting to experimental data sets, yielding higher accuracy and precision than the classic Kok or extended Kok models. This new tool provides a general mathematical framework for analyzing damped oscillations arising from any cycle period using any appropriate Markov model, regardless of symmetry. We illustrate applications of STEAMM that better describe the intrinsic inefficiencies for photon-to-charge conversion within PSII-WOCs that are responsible for damped period-four and period-two oscillations of flash O2 yields across diverse species, while using simpler Markov models free from unrealistic assumptions

NH\u3csub\u3e3\u3c/sub\u3e Binding to the S\u3csub\u3e2\u3c/sub\u3e State of the O\u3csub\u3e2\u3c/sub\u3e-Evolving Complex of Photosystem II: Analogue to H\u3csub\u3e2\u3c/sub\u3eO Binding during the S\u3csub\u3e2\u3c/sub\u3e → S\u3csub\u3e3\u3c/sub\u3e Transition

© 2015 American Chemical Society. Ammonia binds directly to the oxygen-evolving complex of photosystem II (PSII) upon formation of the S2 intermediate, as evidenced by electron paramagnetic resonance spectroscopy. We explore the binding mode by using quantum mechanics/molecular mechanics methods and simulations of extended X-ray absorption fine structure spectra. We find that NH3 binds as an additional terminal ligand to the dangling Mn4, instead of exchanging with terminal water. Because water and ammonia are electronic and structural analogues, these findings suggest that water binds analogously during the S2 → S3 transition, leading to rearrangement of ligands in a carrousel around Mn4

Slow Equilibration between Spectroscopically Distinct Trap States in Reduced TiO\u3csub\u3e2\u3c/sub\u3e Nanoparticles

© 2017 American Chemical Society. Understanding the nature of charge carriers in nanoscale titanium dioxide is important for its use in solar energy conversion, photocatalysis, and other applications. UV-irradiation of aqueous, colloidal TiO2 nanoparticles in the presence of methanol gives highly reduced suspensions. Two distinct types of electron traps were observed and characterized by EPR and optical spectroscopies. The relative populations of the states depend on temperature, indicating a small energy difference, ΔH° = 3.0 ± 0.6 kcal/mol (130 ± 30 meV). Interconversion between the electron traps occurs slowly over the course of minutes to hours within the temperature range studied here, 0-50 °C. The slow time scale implies that interconversion involves changes in structure or stoichiometry, not just the movement of electrons. This occurrence of slow structural modification with changes in trap state occupancy is likely a general feature of reduced TiO2 systems at thermodynamic equilibria or photostationary states and should be considered in the design of TiO2-containing devices

Identification of an oxygenic reaction center psbadc operon in the cyanobacterium gloeobacter violaceus PCC 7421

Gloeobacter violaceus, the earliest diverging oxyphotobacterium (cyanobacterium) on the 16S ribosomal RNA tree, has five copies of the photosystem II psbA gene encoding the D1 reaction center protein subunit. These copies are widely distributed throughout the 4.6 Mbp genome with only one copy colocalizing with other PSII subunits, in marked contrast to all other psbA genes in all publicly available sequenced genomes. A clustering of two other psb genes around psbA3 (glr2322) is unique to Gloeobacter. We provide experimental proof for the transcription of a psbA3DC operon, encoding three of the five reaction center core subunits (D1, D2, and CP43). This is the first example of a transcribed gene cluster containing the D1/D2 or D1/D2/CP43 subunits of PSII in an oxygenic phototroph (prokaryotic or eukaryotic). Implications for the evolution of oxygenic photosynthesis are discussed. © The Author 2011