Access to the Antenna System of Photosystem I via Single-Molecule Excitation-Emission Spectroscopy.

In the development of single-molecule spectroscopy, the simultaneous detection of the excitation and emission spectra has been limited. The fluorescence excitation spectrum based on background-free signals is compatible with the fluorescence-emission-based detection of single molecules and can provide insight into the variations in the input energy of the different terminal emitters. Here, we implement single-molecule excitation-emission spectroscopy (SMEES) for photosystem I (PSI) via a cryogenic optical microscope. To this end, we extended our line-focus-based excitation-spectral microscope system to the cryogenic temperature-compatible version. PSI is one of the two photosystems embedded in the thylakoid membrane in oxygen-free photosynthetic organisms. PSI plays an essential role in electron transfer in the photosynthesis reaction. PSIs of many organisms contain a few red-shifted chlorophylls (Chls) with much lower excitation energies than ordinary antenna Chls. The fluorescence emission spectrum originates primarily from the red-shifted Chls, whereas the excitation spectrum is sensitive to the antenna Chls that are upstream of red-shifted Chls. Using SMEES, we obtained the inclining two-dimensional excitation-emission matrix (2D-EEM) of PSI particles isolated from a cyanobacterium, Thermosynechococcus vestitus (equivalent to elongatus), at about 80 K. Interestingly, by decomposing the inclining 2D-EEMs within time course observation, we found prominent variations in the excitation spectra of the red-shifted Chl pools with different emission wavelengths, strongly indicating the variable excitation energy transfer (EET) pathway from the antenna to the terminal emitting pools. SMEES helps us to directly gain information about the antenna system, which is fundamental to depicting the EET within pigment-protein complexes.

Mechanism of Proton Transfer through the D1-E65/D2-E312 Gate during Photosynthetic Water Oxidation.

In photosystem II, the D1-E65/D2-E312 dyad in the Cl-1 channel has been proposed to play a pivotal role in proton transfer during water oxidation. However, the precise mechanism remains elusive. Here, the proton transfer mechanism within the Cl-1 channel was investigated using quantum mechanics/molecular mechanics calculations. The molecular vibration of the E65/E312 dyad and its deuteration effect revealed that the recently suggested stepwise proton transfer, i.e., initial proton release from the dyad followed by slow reprotonation, does not occur in the Cl-1 channel. Instead, proton transfer is proposed to take place via a conformational change at the E65/E312 dyad, acting as a gate. In its closed form, a proton is trapped within the dyad, preventing forward proton transfer. This closed form converts into the open form, where protonated D1-E65 provides a hydrogen bond to the water network, thereby facilitating fast Grotthuss-type proton transfer.

Overall reaction mechanism of photocatalytic CO2 reduction on a Re(i)-complex catalyst unit of a Ru(ii)-Re(i) supramolecular photocatalyst.

Rhenium(i) complexes fac-[ReI(diimine)(CO)3(L)]n+ are mostly used and evaluated as photocatalysts and catalysts in both photochemical and electrochemical systems for CO2 reduction. However, the selective reduction mechanism of CO2 to CO is unclear, although numerous mechanistic studies have been reported. A Ru(ii)-Re(i) supramolecular photocatalyst with fac-[ReI(diimine)(CO)3{OC(O)OCH2CH2NR2}] (R = C2H4OH) as a catalyst unit (RuC2Re) exhibits very high efficiency, selectivity, and durability of CO formation in photocatalytic CO2 reduction reactions. In this work, the reaction mechanism of photocatalytic CO2 reduction using RuC2Re is fully clarified. Time-resolved IR (TR-IR) measurements using rapid-scan FT-IR spectroscopy with laser flash photolysis verify the formation of RuC2Re(COOH) with a carboxylic acid unit, i.e., fac-[ReI(diimine)(CO)3(COOH)], in the photocatalytic reaction solution. Additionally, this important intermediate is detected in an actual photocatalytic reaction using steady state irradiation. Kinetics analysis of the TR-IR spectra and DFT calculations demonstrated the reaction mechanism of the conversion of the one-electron reduced species of RuC2Re with a fac-[ReI(diimine˙-)(CO)3{OC(O)OCH2CH2NR2}]- unit, which was produced via the photochemical reduction of RuC2Re by 1,3-dimethyl-2-phenyl-2,3-dihydro-1H-benzo[d]imidazole (BIH), to RuC2Re(COOH). The kinetics of the recovery processes of the starting complex RuC2Re from RuC2Re(COOH) accompanying the release of CO and OH- was also clarified. As a side reaction of RuC2Re(COOH), a long-lived carboxylate-ester complex with a fac-[ReI(diimine)(CO)3(COOC2H4NR2)] unit, which was produced by the nucleophilic attack of TEOA to one of the carbonyl ligands of RuC2Re(CO) with a fac-[ReI(diimine)(CO)4]+ unit, was formed during the photocatalytic reaction. This complex works not only as a precursor in another minor CO formation process but also as an external photosensitiser that photochemically reduces the other complexes i.e., RuC2Re, RuC2Re(COOH), and the intermediate that is reductively converted to RuC2Re(COOH).

Absorption changes in Photosystem II in the Soret band region upon the formation of the chlorophyll cation radical [PD1PD2].

Flash-induced absorption changes in the Soret region arising from the [PD1PD2]+ state, the chlorophyll cation radical formed upon light excitation of Photosystem II (PSII), were measured in Mn-depleted PSII cores at pH 8.6. Under these conditions, TyrD is i) reduced before the first flash, and ii) oxidized before subsequent flashes. In wild-type PSII, when TyrD● is present, an additional signal in the [PD1PD2]+-minus-[PD1PD2] difference spectrum was observed when compared to the first flash when TyrD is not oxidized. The additional feature was "W-shaped" with troughs at 434 nm and 446 nm. This feature was absent when TyrD was reduced, but was present (i) when TyrD was physically absent (and replaced by phenylalanine) or (ii) when its H-bonding histidine (D2-His189) was physically absent (replaced by a Leucine). Thus, the simple difference spectrum without the double trough feature at 434 nm and 446 nm, seemed to require the native structural environment around the reduced TyrD and its H bonding partners to be present. We found no evidence of involvement of PD1, ChlD1, PheD1, PheD2, TyrZ, and the Cytb559 heme in the W-shaped difference spectrum. However, the use of a mutant of the PD2 axial His ligand, the D2-His197Ala, shows that the PD2 environment seems involved in the formation of "W-shaped" signal.

Rapid-Scan Fourier Transform Infrared Monitoring of the Photoactivation Process in Cyanobacterial Photosystem II.

The catalytic site of photosynthetic water oxidation, the Mn4CaO5 cluster, in photosystem II (PSII) is known to be formed by a light-induced process called photoactivation. However, details of its molecular mechanism remain unresolved. In this study, we monitored the photoactivation process in cyanobacterial PSII using rapid-scan, time-resolved Fourier transform infrared (FTIR) spectroscopy. The Mn3+/Mn2+ FTIR difference spectra of PSII, in which D1-D170 was specifically 13C labeled, and PSII from the D1-D170A, D1-E189A, and D1-D342A mutants provide strong evidence that the initial Mn2+ is coordinated by D1-D170 and D1-E189. Protein conformational changes and relocation of photo-oxidized Mn3+ in the dark rearrangement process were detected as slow-phase signals in the amide I and carboxylate regions, whereas similar signals were not observed in D1-E189A PSII. It is thus proposed that relocation of Mn3+ via D1-E189 induces the conformational changes of the proteins to form proper Mn binding sites in the mature protein conformation.

Triplet Delocalization over the Reaction Center Chlorophylls in Photosystem II.

The triplet state of chlorophyll formed by charge recombination in photosystem II (PSII) is a precursor of harmful singlet oxygen. Although main localization of the triplet state on the monomeric chlorophyll, ChlD1, at cryogenic temperatures has been suggested, how the triplet state is delocalized on other chlorophylls remains unclear. Here, we investigated the distribution of the triplet state of chlorophyll in PSII using light-induced Fourier transform infrared (FTIR) difference spectroscopy. Measurements of triplet-minus-singlet FTIR difference spectra with PSII core complexes from cyanobacterial mutants, D1-V157H, D2-V156H, D2-H197A, and D1-H198A, in which the interactions of the 131-keto C═O groups of the reaction center chlorophylls, PD1, PD2, ChlD1, and ChlD2, respectively, were perturbed, identified the 131-keto C═O bands of the individual chlorophylls and showed that the triplet state is delocalized over all of these chlorophylls. It is suggested that the triplet delocalization plays important roles in the photoprotection and photodamage mechanisms in PSII.

Role of D1-Glu65 in Proton Transfer during Photosynthetic Water Oxidation in Photosystem II.

Photosynthetic water oxidation takes place at the Mn4CaO5 cluster in photosystem II (PSII) through a light-driven cycle of five intermediates called S states (S0-S4). Although the PSII structures have shown the presence of several channels around the Mn4CaO5 cluster leading to the lumen, the pathways for proton release in the individual S-state transitions remain unidentified. Here, we studied the involvement of the so-called Cl channel in proton transfer during water oxidation by examining the effect of the mutation of D1-Glu65, a key residue in this channel, to Ala using Fourier transform infrared difference and time-resolved infrared spectroscopies together with thermoluminescence and delayed luminescence measurements. It was shown that the structure and the redox property of the catalytic site were little affected by the D1-Glu65Ala mutation. In the S2 → S3 transition, the efficiency was still high and the transition rate was only moderately retarded in the D1-Glu65Ala mutant. In contrast, the S3 → S0 transition was significantly inhibited by this mutation. These results suggest that proton transfer in the S2 → S3 transition occurs through multiple pathways including the Cl channel, whereas this channel likely serves as a single pathway for proton exit in the S3 → S0 transition.

Post-translational amino acid conversion in photosystem II as a possible origin of photosynthetic oxygen evolution.

Photosynthetic oxygen evolution is performed at the Mn cluster in photosystem II (PSII). The advent of this reaction on ancient Earth changed its environment by generating an oxygenic atmosphere. However, how oxygen evolution originated during the PSII evolution remains unknown. Here, we characterize the site-directed mutants at the carboxylate ligands to the Mn cluster in cyanobacterial PSII. A His residue replaced for D1-D170 is found to be post-translationally converted to the original Asp to recover oxygen evolution. Gln/Asn residues in the mutants at D1-E189/D1-D342 are also converted to Glu/Asp, suggesting that amino-acid conversion is a common phenomenon at the ligand sites of the Mn cluster. We hypothesize that post-translational generation of carboxylate ligands in ancestral PSII could have led to the formation of a primitive form of the Mn cluster capable of partial water oxidation, which could have played a crucial role in the evolutionary process of photosynthetic oxygen evolution.

pH-Dependent Regulation of Electron Flow in Photosystem II by a Histidine Residue at the Stromal Surface.

In photosystem II (PSII), the secondary plastoquinone electron acceptor QB functions as a substrate that converts into plastoquinol upon its double reduction by electrons abstracted from water. It has been suggested that a histidine residue, D1-H252, which is located at the stromal surface near QB, is involved in the pH-dependent regulation of electron flow and proton transfer to QB. However, definitive evidence for the involvement of D1-H252 in the QB reactions has not been obtained yet. Here, we studied the roles of D1-H252 in PSII using a cyanobacterial mutant, in which D1-H252 was replaced with Ala. Delayed luminescence (DL) measurement upon a single flash showed a faster QB- decay at higher pH in the thylakoids from the wild-type strain due to the downshift of the redox potential of QB [Em(QB-/QB)]. This pH dependence of the QB- decay was lost in the D1-H252A mutant. The experimental Em(QB-/QB) changes were well reproduced by the density functional theory calculations for models with different protonation states of D1-H252 and with Ala replaced for H252. It was further shown that the period-four oscillation of the DL intensity by successive flashes was significantly diminished in the D1-H252A mutant, suggesting the inhibition of plastoquinone exchange at the QB pocket in this mutant. It is thus concluded that D1-H252 is a key amino acid residue that regulates electron flow in PSII by sensing pH in the stroma and stabilizes the QB binding site to facilitate the quinone exchange reaction.

Redox properties and regulatory mechanism of the iron-quinone electron acceptor in photosystem II as revealed by FTIR spectroelectrochemistry.

Photosystem II (PSII) performs oxidation of water and reduction of plastoquinone through light-induced electron transfer. Electron transfer reactions at individual redox cofactors are controlled by their redox potentials, and the forward and backward electron flows in PSII are regulated by tuning them. It is, thus, crucial to accurately estimate the redox potentials of the cofactors and their shifts by environmental changes to understand the regulatory mechanisms in PSII. Fourier-transform infrared (FTIR) spectroelectrochemistry combined with a light-induced difference technique is a powerful method to investigate the mechanisms of the redox reactions in PSII. In this review, we introduce the methodology and the application of this method in the studies of the iron-quinone complex, which consists of two plastoquinone molecules, QA and QB, and the non-heme iron, on the electron-acceptor side of PSII. It is shown that FTIR spectroelectrochemistry is a useful method not only for estimating the redox potentials but also for detecting the reactions of nearby amino-acid residues coupled with the redox reactions.

D139N mutation of PsbP enhances the oxygen-evolving activity of photosystem II through stabilized binding of a chloride ion.

Photosystem II (PSII) is a multisubunit membrane protein complex that catalyzes light-driven oxidation of water to molecular oxygen. The chloride ion (Cl-) has long been known as an essential cofactor for oxygen evolution by PSII, and two Cl- ions (Cl-1 and Cl-2) have been found to specifically bind near the Mn4CaO5 cluster within the oxygen-evolving center (OEC). However, despite intensive studies on these Cl- ions, little is known about the function of Cl-2, the Cl- ion that is associated with the backbone nitrogens of D1-Asn338, D1-Phe339, and CP43-Glu354. In green plant PSII, the membrane extrinsic subunits-PsbP and PsbQ-are responsible for Cl- retention within the OEC. The Loop 4 region of PsbP, consisting of highly conserved residues Thr135-Gly142, is inserted close to Cl-2, but its importance has not been examined to date. Here, we investigated the importance of PsbP-Loop 4 using spinach PSII membranes reconstituted with spinach PsbP proteins harboring mutations in this region. Mutations in PsbP-Loop 4 had remarkable effects on the rate of oxygen evolution by PSII. Moreover, we found that a specific mutation, PsbP-D139N, significantly enhances the oxygen-evolving activity in the absence of PsbQ, but not significantly in its presence. The D139N mutation increased the Cl- retention ability of PsbP and induced a unique structural change in the OEC, as indicated by light-induced Fourier transform infrared (FTIR) difference spectroscopy and theoretical calculations. Our findings provide insight into the functional significance of Cl-2 in the water-oxidizing reaction of PSII.

Effects of Stromal and Lumenal Side Perturbations on the Redox Potential of the Primary Quinone Electron Acceptor QA in Photosystem II.

The primary quinone electron acceptor QA is a key component in the electron transfer regulation in photosystem II (PSII), and hence accurate estimation of its redox potential, Em(QA-/QA), is crucial in understanding the regulatory mechanism. Although fluorescence detection has been extensively used for monitoring the redox state of QA, it was recently suggested that this method tends to provide a higher Em(QA-/QA) estimate depending on the sample status due to the effect of measuring light [Kato et al. (2019) Biochim. Biophys. Acta 1860, 148082]. In this study, we applied the Fourier transform infrared (FTIR) spectroelectrochemistry, which uses non-reactive infrared light to monitor the redox state of QA, to investigate the effects of stromal- and lumenal-side perturbations on Em(QA-/QA) in PSII. It was shown that replacement of bicarbonate bound to the non-heme iron with formate upshifted Em(QA-/QA) by ∼55 mV, consistent with the previous fluorescence measurement. In contrast, an Em(QA-/QA) difference between binding of 3-(3,4-dichlorophenyl)-1,1-dimethylurea and bromoxynil was found to be ∼30 mV, which is much smaller than the previous estimate, ∼100 mV, by the fluorescence method. This ∼30 mV difference was verified by the decay kinetics of the S2QA- recombination. On the lumenal side, Mn depletion hardly affected the Em(QA-/QA), confirming the previous FTIR result. However, removal of the extrinsic proteins by NaCl or CaCl2 wash downshifted the Em(QA-/QA) by 14-20 mV. These results suggest that electron flow through QA is regulated by changes both on the stromal and lumenal sides of PSII.

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.

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.

ATR-FTIR Spectroelectrochemical Study on the Mechanism of the pH Dependence of the Redox Potential of the Non-Heme Iron in Photosystem II.

The non-heme iron that bridges the two plastoquinone electron acceptors, QA and QB, in photosystem II (PSII) is known to have a redox potential (Em) of ∼+400 mV with a pH dependence of ∼-60 mV/pH. However, titratable amino acid residues that are coupled to the redox reaction of the non-heme ion and responsible for its pH dependence remain unidentified. In this study, to clarify the mechanism of the pH dependent change of Em(Fe2+/Fe3+), we investigated the protonation structures of amino acid residues correlated with the pH-induced Em(Fe2+/Fe3+) changes using Fourier transform infrared (FTIR) spectroelectrochemistry combined with the attenuated total reflection (ATR) and light-induced difference techniques. Flash-induced Fe2+/Fe3+ ATR-FTIR difference spectra obtained at different electrode potentials in the pH range of 5.0-8.5 showed a linear pH dependence of Em(Fe2+/Fe3+) with a slope of -52 mV/pH close to the theoretical value at 10 °C, the measurement temperature. The spectral features revealed that D1-H215, a ligand to the non-heme iron interacting with QB, was deprotonated to an imidazolate anion at higher pH with a pKa of ∼5.6 in the Fe3+ state, while carboxylate groups from Glu/Asp residues present on the stromal side of PSII were protonated at lower pH with a pKa of ∼5.7 in the Fe2+ state. It is thus concluded that the deprotonation/protonation reactions of D1-H215 and Glu/Asp residues located near the non-heme iron cause the pH-dependent changes in Em(Fe2+/Fe3+) at higher and lower pH regions, respectively, realizing a linear pH dependence over a wide pH range.

Rapid-Scan Time-Resolved ATR-FTIR Study on the Photoassembly of the Water-Oxidizing Mn4CaO5 Cluster in Photosystem II.

The catalytic center of photosynthetic water oxidation, the Mn4CaO5 cluster, is assembled in photosystem II (PSII) through a light-driven process called photoactivation, whose mechanism remains elusive. Here, we used rapid-scan time-resolved Fourier transform infrared (FTIR) spectroscopy combined with the attenuated total reflection (ATR) technique to monitor the photoactivation process. Rapid-scan ATR-FTIR spectra of apo-PSII with Mn2+ upon flash illumination showed spectral features typical of carboxylate stretching vibrations, which were attributed to two carboxylate ligands, D1-D170 and D1-E189, by quantum chemical calculations. The FTIR signal decayed with a time constant of ∼0.7 s, showing that the subsequent "dark rearrangement" step occurred with a low quantum yield and Mn3+ ions were mostly released during this decay. Simulation of the kinetic process provided a slow intrinsic rate of the dark rearrangement, which was attributed to a large protein conformational change. The photoassembly mechanism of the Mn4CaO5 cluster is proposed based on these findings.

Protonation State of a Key Histidine Ligand in the Iron-Quinone Complex of Photosystem II as Revealed by Light-Induced ATR-FTIR Spectroscopy.

The iron-quinone complex in photosystem II (PSII) consists of the two plastoquinone electron acceptors, QA and QB, and a non-heme iron connecting them. It has been suggested that nearby histidine residues play important roles in the electron and proton transfer reactions of the iron-quinone complex in PSII. In this study, we investigated the protonation/deprotonation reaction of D1-H215, which bridges the non-heme iron and QB, using attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy. Flash-induced Fe2+/Fe3+ ATR-FTIR difference spectra were measured with PSII membranes in the pH range of 5.0-7.5. In the CN stretching region of histidine, the intensity of a negative peak at 1094 cm-1, which was assigned to the deprotonated anion form of D1-H215, increased as the pH increased. Singular-value decomposition analysis provided a component due to deprotonation of D1-H215 with a pKa of ∼5.5 in the Fe3+ state, whereas no component of histidine deprotonation was resolved in the Fe2+ state. This observation supports the previous proposal that D1-H215 is responsible for the proton release upon Fe2+ oxidation [Berthomieu, C., and Hienerwadel, R. (2001) Biochemistry 40, 4044-4052]. The pH dependence of the 13C isotope-edited bands of the bicarbonate ligand to the non-heme iron further showed that deprotonation of bicarbonate to carbonate does not take place at pH <8 in the Fe2+ or Fe3+ state. These results suggest that the putative mechanism of proton transfer to QBH- through D1-H215 and bicarbonate around Fe2+ functions throughout the physiological pH range.

Protonation structure of the photosynthetic water oxidizing complex in the S0 state as revealed by normal mode analysis using quantum mechanics/molecular mechanics calculations.

Photosynthetic water oxidation takes place through the light-driven cycle of five intermediates (S0-S4) of the water oxidizing complex (WOC), which consists of the Mn4CaO5 cluster and surrounding amino acid residues in photosystem II. Clarifying the protonation structures of the Mn4CaO5 cluster and its water ligands (W1-W4) is essential for understanding the molecular mechanism of water oxidation. Here, we performed normal mode analysis of WOC in the S0 and S1 states using quantum mechanics/molecular mechanics calculations and simulated an S1-minus-S0 infrared difference spectrum focusing on the symmetric COO- stretching (νsCOO-) region. The calculated spectrum by an S0 model, in which O4 of the Mn4CaO5 cluster is protonated and W2 is H2O, and a corresponding S1 state with deprotonated O4 best reproduced the νsCOO- features of the experimental spectrum, whereas models with protonated O5 showed poor agreement. In addition, comparison of the calculated coordination distances of the water ligands with the experimental data by X-ray diffraction analysis indicates that W2 is most probably not OH- but H2O both in the S0 and S1 states. The present calculations thus strongly suggest that the S0 state has a protonation structure of O4-H and W2 = H2O. The O4-H structure in the S0 state supports the view that this proton is released through the O4-water chain immediately after electron transfer during the S0→ S1 transition.

Formation of the High-Spin S2 State Related to the Extrinsic Proteins in the Oxygen Evolving Complex of Photosystem II.

The high-spin S2 state was investigated with photosystem II (PSII) from spinach, Thermosynechococcus vulcanus, and Cyanidioschyzon merolae. In extrinsic protein-depleted PSII, high-spin electron paramagnetic resonance (EPR) signals were not detected in either species, whereas all species showed g ∼ 5 signals in the presence of a high concentration of Ca2+ instead of the multiline signal. In the intact and PsbP/Q-depleted PSII from spinach, the g = 4.1 EPR signal was detected. These results show that formation of the high-spin S2 state of the manganese cluster is regulated by the extrinsic proteins through a charge located near the Mn4 atom in the Mn4CaO5 cluster but is independent of the intrinsic proteins. The shift to the g ∼ 5 state is caused by tilting of the z-axis in the Mn4 coordinates through hydrogen bonds or external divalent cations. The structural modification may allow insertion of an oxygen atom during the S2-to-S3 transition.

Structural Dynamics of a Protein Domain Relevant to the Water-Oxidizing Complex in Photosystem II as Visualized by High-Speed Atomic Force Microscopy.

Photosystem II (PSII) is a multiprotein complex that has a function of light-driven water oxidation. The catalytic site of water oxidation is the Mn4CaO5 cluster, which is bound to the lumenal side of PSII through amino acid residues from the D1 and CP43 proteins and is further surrounded by the extrinsic proteins. In this study, we have for the first time visualized the structural dynamics of the lumenal region of a PSII core complex using high-speed atomic force microscopy (HS-AFM). The HS-AFM images of a PSII membrane fragment showed stepwise dissociation of the PsbP and PsbO extrinsic proteins. Upon subsequent destruction of the Mn4CaO5 cluster, the lumenal domain of CP43 was found to undergo a conformational fluctuation. The observed structural flexibility and conformational fluctuation of the CP43 lumenal domain are suggested to play important roles in the biogenesis of PSII and the photoassembly of the Mn4CaO5 cluster.

Distribution Map of Frost Resistance for Cement-Based Materials Based on Pore Structure Change.

This paper presents a prediction method and mathematical model based on experimental results for the change in pore structure of cement-based materials due to environmental conditions. It focuses on frost damage risk to cement-based materials such as mortar. Mortar specimens are prepared using water, ordinary Portland cement, and sand and the pore structure is evaluated using mercury intrusion porosimetry. New formulas are proposed to describe the relationship between the pore structure change and the modified maturity and to predict the durability factor. A quantitative prediction model is established from a modified maturity function considering the influences of environmental factors like temperature and relative humidity. With this model, the frost resistance of cement-based materials can be predicted based on weather data. Using the prediction model and climate data, a new distribution map of frost damage risk is created. It is found that summer weather significantly affects frost resistance, owing to the change in pore structure of cement-based mortar. The model provides a valuable tool for predicting frost damage risk based on weather data and is significant for further research.