Role Of Ca2 Research Articles

Pollen tube in hazel grows intermittently: Role of Ca2+ and expression of auto-inhibited Ca2+ pump

Owing to the absence of the ovary when the female inflorescence blooms, hazel pollen tubes show an obvious intermittent growth pattern, which has an important impact on the number of fruits per cluster and yield, but little is known about the mechanisms governing this process. Spray treatments of Ca2+ and Ca2+-ATPase activator 5-aminolevulinic acid (ALA) in field experiment showed that Ca2+ treatments inhibited pistillate inflorescence drop and produced more fruits, while the effects of ALA treatments was just opposite to that of calcium treatments. Highly accumulated Ca2+ was observed at the rear part of the stigma and in the ovary primordial cell layers. Under in vitro culture conditions, pollen germination ratio and tube length were promoted and inhibited by Ca2+ and ALA, respectively. ALA treatment reduced the Ca2+ concentration in pollen tube, which was not conducive to the formation of a tip-focused Ca2+ gradient in pollen tube. The hazel genome encoded 17 Ca2+-ATPases, including 14 auto-inhibited Ca2+-ATPases and three ER-type Ca2+-ATPases. Hazel Ca2+-ATPase family proteins, including 11 common transmembrane domains, were highly conserved. Of 17 Ca2+-ATPases, qRT-PCR analysis showed that ChACA1 and ChECA2 were highly expressed in pollen tubes. In the range of 0–1.3 mM Ca2+, Ca2+ fluorescence intensity in pollen tubes, expression of ChACA1 continued to rise simultaneously, and they reached their maximum at 1.3 mM Ca2+, then declined at 1.6 mM Ca2+, which was consistent and inconsistent with changing trends of pollen tube length and ChECA2 respectively. Analysis of enzyme activity in pollen tubes showed that Ca2+-ATPase activity was promoted by both Ca2+ and ALA treatments significantly. Our findings suggested that ChACA1 may act as a key regulator of pollen germination and pollen tube development in hazel, and provide new insight into the mechanisms of unique pollen tube intermittent growth in hazel.

Elucidating the Mechanism of Membrane Destabilization by the Preferred Modes of Insertion of the Sars-Cov2 Fusion Peptide

Cell penetration of the SARS-Cov2 virus after recognition by the ACE2 receptor requires the fusion of the viral envelope membrane with cellular membranes. The Spike protein (S) of the virus harbors a region identified as the “fusion peptide” (FP) which is liberated at its N-terminal site by a specific cleavage occurring in concert with the interaction of the receptor binding domain of the Spike. Studies have shown that the SARS-Cov2-FP binds Ca2+ ions and perturbs membranes in a calcium-dependent fashion. But the mechanisms of membrane insertion and destabilization remained unclear. We have identified the preferred modes of SARS-Cov2-FP insertion and the role of Ca2+ ions in mediating peptide-membrane interactions from extensive atomistic molecular dynamics (MD) simulations and trajectory analyses. Multiple Ca2+ binding modes in which the ion engaged with different pairs of acidic residues on the FP were observed. A systematic sampling of the interactions of these Ca2+-bound peptide models with lipid membranes showed that SARS-Cov2-FP penetrated the bilayer only in two modes involving different structural domains. In the first mode the hydrophobic residues F833/I834 from the middle region of the peptide are inserted. The second, and more prevalent mode of penetration involves residues L822/F823 in the freed N-terminus of the peptide. The latter insertion was related to the specific mode of Ca2+ association with the peptide in which one Ca2+ is bound to the D830/D839 pair and another to E819/D820. These findings explain the mechanistic role of the necessary cleavage (termed S2’) that frees the N-terminus of the peptide to effectuate the cell-entry of SARS-Cov2. Computational resources provided by the COVID-19 HPC Consortium are gratefully acknowledged.

Collapsin response mediator protein4 enhances the radiosensitivity of colon cancer cells through calcium‑mediated cell signaling.

Radiation therapy is an effective treatment against various types of cancer, but some radiation-resistant cancer cells remain a major therapeutic obstacle; thus, understanding radiation resistance mechanisms is essential for cancer treatment. In this study, we established radiation-resistant colon cancer cell lines and examined the radiation-induced genetic changes associated with radiation resistance. Using RNA-sequencing analysis, collapsin response mediator protein 4 (CRMP4) was identified as the candidate gene associated with radiation sensitivity. When cells were exposed to radiation, intracellular Ca2+ influx, collapse of mitochondrial membrane potential, and cytochrome c release into the cytosol were increased, followed by apoptosis induction. Radiation treatment- or Ca2+ ionophore A23187-induced apoptosis was significantly inhibited in CRMP4-deficient cells, including radiation-resistant or CRMP4-shRNA cell lines. Furthermore, treatment of CRMP4-deficient cells with low levels (<5 µM) of BAPTA-AM, a Ca2+ chelator, resulted in radiation resistance. Conversely, Ca2+ deficiency induced by a high BAPTA-AM concentration (>10 µM) resulted in higher cell death in the CRMP4-depleted cells compared to CRMP4-expressing control cells. Our results suggest that CRMP4 plays an important role in Ca2+-mediated cell death pathways under radiation exposure and that CRMP4 may be a therapeutical target for colon cancer treatment.

SERCA2a ameliorates cardiomyocyte T-tubule remodeling via the calpain/JPH2 pathway to improve cardiac function in myocardial ischemia/reperfusion mice

Transverse-tubules (T-tubules) play pivotal roles in Ca2+-induced, Ca2+ release and excitation–contraction coupling in cardiomyocytes. The purpose of this study was to uncover mechanisms where sarco/endoplasmic reticulum Ca2+ ATPase (SERCA2a) improved cardiac function through T-tubule regulation during myocardial ischemia/reperfusion (I/R). SERCA2a protein expression, cytoplasmic [Ca2+]i, calpain activity, junctophilin-2 (JPH2) protein expression and intracellular localization, cardiomyocyte T-tubules, contractility and calcium transients in single cardiomyocytes and in vivo cardiac functions were all examined after SERCA2a knockout and overexpression, and Calpain inhibitor PD150606 (PD) pretreatment, following myocardial I/R. This comprehensive approach was adopted to clarify SERCA2a mechanisms in improving cardiac function in mice. Calpain was activated during myocardial I/R, and led to the proteolytic cleavage of JPH2. This altered the T-tubule network, the contraction function/calcium transients in cardiomyocytes and in vivo cardiac functions. During myocardial I/R, PD pretreatment upregulated JPH2 expression and restored it to its intracellular location, repaired the T-tubule network, and contraction function/calcium transients of cardiomyocytes and cardiac functions in vivo. SERCA2a suppressed calpain activity via [Ca2+]i, and ameliorated these key indices. Our results suggest that SERCA2a ameliorates cardiomyocyte T-tubule remodeling via the calpain/JPH2 pathway, thereby improving cardiac function in myocardial I/R mice.

Intracellular Ca2 + Imbalance Critically Contributes to Paraptosis.

Paraptosis is a type of programmed cell death that is characterized by dilation of the endoplasmic reticulum (ER) and/or mitochondria. Since paraptosis is morphologically and biochemically different from apoptosis, understanding its regulatory mechanisms may provide a novel therapeutic strategy in malignant cancer cells that have proven resistant to conventional pro-apoptotic treatments. Relatively little is known about the molecular basis of paraptosis, but perturbations of cellular proteostasis and ion homeostasis appear to critically contribute to the process. Ca2+ transport has been shown to be important in the paraptosis induced by several natural products, metal complexes, and co-treatment with proteasome inhibitors and certain Ca2+-modulating agents. In particular, the Ca2+-mediated communication between the ER and mitochondria plays a crucial role in paraptosis. Mitochondrial Ca2+ overload from the intracellular Ca2+-flux system located at the ER–mitochondrial axis can induce mitochondrial dilation during paraptosis, while the accumulation of misfolded proteins within the ER lumen is believed to exert an osmotic force and draw water from the cytoplasm to distend the ER lumen. In this process, Ca2+ release from the ER also critically contributes to aggravating ER stress and ER dilation. This review focuses on the role of Ca2+ transport in paraptosis by summarizing the recent findings related to the actions of Ca2+-modulating paraptosis-inducing agents and discussing the potential cancer therapeutic strategies that may effectively induce paraptosis via Ca2+ signaling.

Intracellular Ca2+ Signaling in Protozoan Parasites: An Overview with a Focus on Mitochondria.

Ca2+ signaling has been involved in controling critical cellular functions such as activation of proteases, cell death, and cell cycle control. The endoplasmatic reticulum plays a significant role in Ca2+ storage inside the cell, but mitochondria have long been recognized as a fundamental Ca2+ pool. Protozoan parasites such as Plasmodium falciparum, Toxoplasma gondii, and Trypanosoma cruzi display a Ca2+ signaling toolkit with similarities to higher eukaryotes, including the participation of mitochondria in Ca2+-dependent signaling events. This review summarizes the most recent knowledge in mitochondrial Ca2+ signaling in protozoan parasites, focusing on the mechanism involved in mitochondrial Ca2+ uptake by pathogenic protists.

Ryanodine receptor remodeling in cardiomyopathy and muscular dystrophy caused by lamin A/C gene mutation.

Mutations in the lamin A/C gene (LMNA), which encodes A-type lamins, cause several diseases called laminopathies, the most common of which is dilated cardiomyopathy with muscular dystrophy. The role of Ca2+ regulation in these diseases remain poorly understood. We now show biochemical remodeling of the ryanodine receptor (RyR)/intracellular Ca2+ release channel in heart samples from human subjects with LMNA mutations, including protein kinase A-catalyzed phosphorylation, oxidation and depletion of the stabilizing subunit calstabin. In the LmnaH222P/H222P murine model of Emery-Dreifuss muscular dystrophy caused by LMNA mutation, we demonstrate an age-dependent biochemical remodeling of RyR2 in the heart and RyR1 in skeletal muscle. This RyR remodeling is associated with heart and skeletal muscle dysfunction. Defective heart and muscle function are ameliorated by treatment with a novel Rycal small molecule drug (S107) that fixes 'leaky' RyRs. SMAD3 phosphorylation is increased in hearts and diaphragms of LmnaH222P/H222P mice, which enhances NADPH oxidase binding to RyR channels, contributing to their oxidation. There is also increased generalized protein oxidation, increased calcium/calmodulin-dependent protein kinase II-catalyzed phosphorylation of RyRs and increased protein kinase A activity in these tissues. Our data show that RyR remodeling plays a role in cardiomyopathy and skeletal muscle dysfunction caused by LMNA mutation and identify these Ca2+ channels as a potential therapeutic target.

Advances in the study of cancer metastasis and calcium signaling as potential therapeutic targets.

Metastasis is still the primary cause of cancer-related mortality. However, the underlying mechanisms of cancer metastasis are not yet fully understood. Currently, the epithelial-mesenchymal transition, metabolic remodeling, cancer cell intercommunication and the tumor microenvironment including diverse stromal cells, are reported to affect the metastatic process of cancer cells. Calcium ions (Ca2+) are ubiquitous second messengers that manipulate cancer metastasis by affecting signaling pathways. Diverse transporter/pump/channel-mediated Ca2+ currents form Ca2+ oscillations that can be decoded by Ca2+-binding proteins, which are promising prognostic biomarkers and therapeutic targets of cancer metastasis. This paper presents a review of the advances in research on the mechanisms underlying cancer metastasis and the roles of Ca2+-related signals in these events.

A novel strategy for treating cancer: understanding the role of Ca2+ signaling from nociceptive TRP channels in regulating cancer progression.

Cancer is an aging-associated disease and caused by genomic instability that is driven by the accumulation of mutations and epimutations in the aging process. Although Ca2+ signaling, reactive oxygen species (ROS) accumulation, DNA damage response (DDR) and senescence inflammation response (SIR) are processed during genomic instability, the underlying mechanism for the cause of genomic instability and cancer development is still poorly understood and needs to be investigated. Nociceptive transient receptor potential (TRP) channels, which firstly respond to environmental stimuli, such as microbes, chemicals or physical injuries, potentiate regulation of the aging process by Ca2+ signaling. In this review, the authors provide an explanation of the dual role of nociceptive TRP channels in regulating cancer progression, initiating cancer progression by aging-induced genomic instability, and promoting malignancy by epigenetic regulation. Thus, therapeutically targeting nociceptive TRP channels seems to be a novel strategy for treating cancers.

TSPAN-7 as a key regulator of glucose-stimulated Ca2+ influx and insulin secretion.

TSPAN-7 as a key regulator of glucose-stimulated Ca2+ influx and insulin secretion.

Structural insights into a C2 domain protein specifically found in tardigrades.

Some tardigrades can survive extremely desiccated conditions through transition into a state called anhydrobiosis. Anhydrobiotic tardigrades have proteins unique to them and they are thought to be keys to the understanding of unusual desiccation resistance. In fact, previous transcriptome data show that several tardigrade‐specific proteins are significantly upregulated under desiccated conditions. However, their physiological roles and chemical properties have been ambiguous because they show low or no similarity of amino acid sequences to proteins found in other organisms. Here, we report a crystal structure of one of such proteins. This protein shows a β‐sandwich structure composed of 8 β‐strands, three Ca2+‐binding sites, and hydrophobic residues on Ca2+‐binding (CBD) loops, which resemble characteristics of C2 domain proteins. We therefore conveniently describe this protein as tardigrade C2 domain protein (TC2P). Because the C2 domain functions as a Ca2+‐mediated membrane docking module, which is related to signal transduction or membrane trafficking, TC2Ps may play a role in Ca2+‐triggered phenomenon under desiccated situations. Our finding provides not only structural insights into a newly discovered desiccation‐related protein family but also insights into the evolution and diversity of C2 domain proteins.

Emerging roles of the CBL-CIPK calcium signaling network as key regulatory hub in plant nutrition

Plant physiology and development essentially depend on sufficient uptake of various essential nutritive ions via their roots and their appropriate transport and distribution within the organism. Many of these essential nutrients are heterogeneously distributed in the soil or are available in fluctuating concentrations. This natural situation requires constant regulatory adjustment and balancing of nutrient uptake and homeostasis. Here, we review recent findings on the role of Ca2+ signals and Ca2+-dependent regulation via the CBL-CIPK Ca2+ sensor-protein kinase network in these processes. We put special emphasis on Ca2+ controlled processes that contribute to establishing the homeostasis of macro-nutrients like potassium (K+), nitrogen (N), and magnesium (Mg2+) and on the micro-nutrient iron (Fe). Increasing experimental evidence indicates the occurrence of nutrient-specific, spatially and temporally defined cytoplasmic Ca2+ elevations as early responses to nutrient fluctuations. Specific CBL-CIPK complexes translate these signals into phosphorylation regulation of important channels and transporters like AKT1, NPF6.3/NRT1.1, AMT1, SLAC1, TPK1 and IRT1. We discuss a crucial and coordinating role for these Ca2+ signaling mechanisms in regulating the sensing, uptake, distribution and storage of various ions. Finally, we reflect on the emerging multifaceted and potentially integrating role of the “nutrient” kinase CIPK23 in regulating multiple nutrient responses. From this inventory, we finally deduce potential mechanisms that can convey the coordinated regulation of distinct steps in the transport of one individual ion and mechanisms that can bring about the integration of adaptive responses to fluctuations of different ions to establish a faithfully balanced plant nutrient homeostasis.

Thrombin Stimulation of Human Endothelial Cell Phospholipase D Activity. Regulation by Phospholipase C, Protein Kinase C, and Cyclic Adenosine 3' 5’ -Monophosphate

The activation of membrane-bound phospholipase D (PLD) resulting in the generation of phosphatidic acid (PA) is increasingly recognized as an integral event in the initiation of a variety of cellular responses. We explored whether alpha-thrombin is a physiologic agonist for PLD activation in human umbilical vein endothelial cells (HUVEC). HUVEC monolayers were labeled with [32Pi] and PLD activity determined by formation of the PLD metabolite [32P] phosphatidylethanol (PEt) in the presence of 5 g/L ethanol by thin-layer chromatography. alpha-Thrombin rapidly (1 minute) increased PA and PEt formation in a dose-dependent manner (10(-6) to 10(-10)) with maximal PLD stimulation achieved with 10 nmol/L alpha-thrombin producing a threefold to fourfold increase in PA and a sixfold to eightfold increase in PEt over controls at 15 minutes. Esterolytically active zeta-thrombin (10 nmol/L) and gamma-thrombin (1 mumol/L), but not inactive DIP-alpha-thrombin (1 mumol/L) also increased PLD activity. The role of Ca2+ flux in human endothelial cell PLD activation was investigated and PEt formation was significantly enhanced by Ca2+ ionophores A23187 and ionomycin (1 mumol/L, three-fold to fourfold increase in PEt). Alpha-Thrombin-stimulated PEt formation was abolished (greater than 90% inhibition) with chelation of intracellular calcium (Ca2+i) by pretreatment with BAPTA-AM (25 mumol/L, 30 minutes) but only mildly attenuated (30% inhibition) by removal of extracellular calcium (Ca2+E) with EGTA (5 mmol/L). The protein kinase C (PKC) inhibitor staurosporine reduced alpha-thrombin-induced PEt formation in a dose-dependent manner (10 mumol/L, 78% inhibition) and PKC downregulation with chronic PMA treatment (18 hours) also resulted in marked inhibition of alpha-thrombin-induced PEt formation. Neither pertussis nor botulinum C bacterial toxins significantly altered alpha-thrombin-induced PLD responses. In contrast, similar pretreatment with cholera toxin (1 microgram/mL, 60 minutes) consistently augmented alpha-thrombin-stimulated PLD activity by 50% to 90%. Comparable results were observed with agents which increased cAMP such as forskolin, 8-bromo cAMP, or dibutyryl cAMP and cholera toxin augmentation was abolished by 2-dideoxyadenosine, a competitive inhibitor of adenylyl cyclase activity. These studies demonstrate that alpha-thrombin is a potent stimulus for human PLD-mediated PA formation and that cyclic adenosine nucleotides modulate agonist-induced cellular PLD activity. In this model of PLD activation, alpha-thrombin receptor occupancy leads to the breakdown of phosphatidylinositol 4,5-bisphosphate catalyzed by phospholipase C producing the Ca2+ secretagogue IP3 and DAG.(ABSTRACT TRUNCATED AT 400 WORDS)

CarP, encoding a Ca2+-regulated putative phytase, is evolutionarily conserved in Pseudomonas aeruginosa and has potential as a biomarker.

Pseudomonas aeruginosa infects patients with cystic fibrosis, burns, wounds and implants. Previously, our group showed that elevated Ca2+ positively regulates the production of several virulence factors in P. aeruginosa, such as biofilm formation, production of pyocyanin and secreted proteases. We have identified a Ca2+-regulated β-propeller putative phytase, CarP, which is required for Ca2+ tolerance, regulation of the intracellular Ca2+ levels, and plays a role in Ca2+ regulation of P. aeruginosa virulence. Here, we studied the conservation of carP sequence and its occurrence in diverse phylogenetic groups of bacteria. In silico analysis revealed that carP and its two paralogues PA2017 and PA0319 are primarily present in P. aeruginosa and belong to the core genome of the species. We identified 155 single nucleotide alterations within carP, 42 of which lead to missense mutations with only three that affected the predicted 3D structure of the protein. PCR analyses with carP-specific primers detected P. aeruginosa specifically in 70 clinical and environmental samples. Sequence comparison demonstrated that carP is overall highly conserved in P. aeruginosa isolated from diverse environments. Such evolutionary preservation of carP illustrates its importance for P. aeruginosa adaptations to diverse environments and demonstrates its potential as a biomarker.

Calcium-hydrogen sulfide crosstalk during K+-deficient NaCl stress operates through regulation of Na+/H+ antiport and antioxidative defense system in mung bean roots

Present investigation reports the role of calcium (Ca2+) and hydrogen sulfide (H2S) crosstalk associated with Vigna radiata seedlings subjected to K+ deficient conditions under short-term (24 h) and long-term (72 h) NaCl stress. Perusal of the data reveals that under short-term NaCl stress an initial decline in K+ level led to the elevation in Ca2+ and H2S levels along with improvement in antioxidant system and reduction in reactive oxygen species (ROS) production. Under long-term NaCl stress a further decline in K+ content was deleterious that led to a lower K+/Na+ ratio. This was followed by reduction in antioxidant system along with excessive accumulation of ROS and methylglyoxal content, and increased membrane damage. However, supplementation of the seedling roots with Ca2+ enhanced biosynthesis of H2S through enhancing cysteine pool. The present findings suggest that synergistic action of Ca2+ and H2S induced the activity of H+-ATPase that created H+ gradient which in turn induced Na+/H+ antiport system that accelerated K+ influx and Na+ efflux. All of these together contributed to a higher K+/Na+ ratio, activation of antioxidative defense system, and maintenance of redox homeostasis and membrane integrity in Ca2+-supplemented stressed seedlings. Role of Ca2+ and H2S in the regulation of Na+/H+ antiport system was validated by the use of sodium orthovanadate (plasma membrane H+-ATPase inhibitor), tetraethylammonium chloride (K+ channel blocker), and amiloride (Na+/H+ antiporter inhibitor). Application of Ca2+-chelator EGTA (ethylene glycol-bis(b-aminoethylether)-N,N,Nʹ,Nʹ-tetraacetic acid) and H2S scavenger hypotaurine abolished the effect of Ca2+, suggesting the involvement of Ca2+ and H2S in the alleviation of NaCl stress. Moreover, use of EGTA and HT also substantiates the downstream functioning of H2S during Ca2+-mediated regulation of plant adaptive responses to NaCl stress. To sum up, present findings reveal the association of Ca2+ and H2S signaling in the regulation of ion homeostasis and antioxidant defense during K+-deficient NaCl stress.

Bisphenol AF induces apoptosis via estrogen receptor beta (ERβ) and ROS-ASK1-JNK MAPK pathway in human granulosa cell line KGN.

Bisphenol AF (BPAF) is an emerging environmental pollutant. Although BPAF is widely spread in the environment and human surroundings, its interference with ovarian function has not been fully elucidated. The aim of this study was to identify the mechanism underlying the effect of BPAF on the apoptosis of KGN cells, which maintain the physiological characteristics of ovarian granulosa cells. Our results indicated that BPAF induces KGN cell apoptosis in a concentration- and time-dependent manner. Meanwhile, BPAF exposure significantly promoted the expression of pro-apoptotic proteins, including Bax, Bid and Bak, while the expression of anti-apoptotic proteins, such as Bcl-2, Bcl-xL and Mcl-1, decreased significantly. We further detected a significant increase in intracellular ROS levels in response to high concentrations of BPAF exposure. After blocking the corresponding pathway, it was found that ROS mediates ASK1 and JNK activation. Furthermore, the role of Ca2+ overload and estrogen receptor β (ERβ) in BPAF-induced KGN cell apoptosis was also confirmed by using inhibitors. These results suggest that BPAF has potential reproductive toxicity for females, and ROS-ASK1-JNK axis may play a key role in BPAF-induced ovarian dysfunction. In addition, Ca2+ overload and ERβ pathway activation may also be an important mechanism of reproductive toxicity of BPAF.

Biochemical characterization of four splice variants of mouse Ca2+/calmodulin-dependent protein kinase Iδ.

Ca2+/calmodulin (CaM)-dependent protein kinase Iδ (CaMKIδ) is a Ser/Thr kinase that plays pivotal roles in Ca2+ signalling. CaMKIδ is activated by Ca2+/CaM-binding and phosphorylation at Thr180 by CaMK kinase (CaMKK). In this study, we characterized four splice variants of mouse CaMKIδ (mCaMKIδs: a, b, c and d) found by in silico analysis. Recombinant mCaMKIδs expressed in Escherichia coli were phosphorylated by CaMKK; however, only mCaMKIδ-a and c showed protein kinase activities towards myelin basic protein in vitro, with mCaMKIδ-b and mCaMKIδ-d being inactive. Although mCaMKIδ-a and mCaMKIδ-c underwent autophosphorylation in vitro, only mCaMKIδ-c underwent autophosphorylation in 293T cells. Site-directed mutagenesis showed that the autophosphorylation site is Ser349, which is found in the C-terminal region of only variants c and b (Ser324). Furthermore, phosphorylation of these sites (Ser324 and Ser349) in mCaMKIδ-b and c was more efficiently catalyzed by cAMP-dependent protein kinase in vitro and in cellulo as compared to the autophosphorylation of mCaMKIδ-c. Thus, variants of mCaMKIδ possess distinct properties in terms of kinase activities, autophosphorylation and phosphorylation by another kinase, suggesting that they play physiologically different roles in murine cells.

Roles of Ca2+, Mg2+, and Ba2+ Cations in the Regulation of TRPV1 Channels in Rat DRG Neurons

Roles of Ca2+, Mg2+, and Ba2+ Cations in the Regulation of TRPV1 Channels in Rat DRG Neurons

The role of Ca2+ and protein scaffolding in the formation of nature’s water oxidizing complex

Photosynthetic O2 evolution is catalyzed by the Mn4CaO5 cluster of the water oxidation complex of the photosystem II (PSII) complex. The photooxidative self-assembly of the Mn4CaO5 cluster, termed photoactivation, utilizes the same highly oxidizing species that drive the water oxidation in order to drive the incorporation of Mn2+ into the high-valence Mn4CaO5 cluster. This multistep process proceeds with low quantum efficiency, involves a molecular rearrangement between light-activated steps, and is prone to photoinactivation and misassembly. A sensitive polarographic technique was used to track the assembly process under flash illumination as a function of the constituent Mn2+ and Ca2+ ions in genetically engineered membranes of the cyanobacterium Synechocystis sp. PCC6803 to elucidate the action of Ca2+ and peripheral proteins. We show that the protein scaffolding organizing this process is allosterically modulated by the assembly protein Psb27, which together with Ca2+ stabilizes the intermediates of photoactivation, a feature especially evident at long intervals between photoactivating flashes. The results indicate three critical metal-binding sites: two Mn and one Ca, with occupation of the Ca site by Ca2+ critical for the suppression of photoinactivation. The long-observed competition between Mn2+ and Ca2+ occurs at the second Mn site, and its occupation by competing Ca2+ slows the rearrangement. The relatively low overall quantum efficiency of photoactivation is explained by the requirement of correct occupancy of these metal-binding sites coupled to a slow restructuring of the protein ligation environment, which are jointly necessary for the photooxidative trapping of the first stable assembly intermediate.

On fast LuAG:Ce scintillation ceramics with Ca2+ co‐dopants

Abstract“Defect engineering” was a valid strategy to modify the performance of LuAG:Ce scintillator, usually realized by Me2+/Me+ co‐doping. To investigate the effects of Ca2+ co‐doping on the scintillation properties of LuAG:Ce, a set of LuAG:Ce ceramics with Ca2+ concentrations ranging from 0 to 0.5 at.% were manufactured. The absorption spectra, radioluminescence spectra (RL spectra), light yield, RL spectra as a function of temperature, decay time, and TSL curves of the ceramic products were carefully measured. With Ca2+ co‐doping, the scintillation performance of LuAG:Ce ceramics was greatly improved. Especially for the 0.2 at.% Ca2+ co‐doped one, it has a high light yield value of 24, 400 ph/MeV, a fast scintillation decay time of 48 ns, and a small slow component contamination. And the role of Ca2+ in the scintillation mechanism of LuAG:Ce ceramics was also discussed in this paper.