All‐trans‐retinoic acid intensifies endoplasmic reticulum stress in N‐acetylglucosaminyltransferase V repressed human hepatocarcinoma cells by perturbing homocysteine metabolism

We previously reported that all‐trans‐retinoic acid (ATRA) induced apoptosis in N‐acetylglucosaminyltransferase V (GnT‐V) repressed human hepatocarcinoma 7721 (GnT‐V‐AS/7721) cells via endoplasmic reticulum (ER) stress. In addition to confirming these findings, we further found that ATRA repressed the expression of betaine‐homocysteine methyltransferase (BHMT) and cystathionine‐β‐synthase (CBS), which are key enzymes that are involved in homocysteine metabolism, increased the level of intracellular homocysteine, and decreased the glutathione (GSH) level in GnT‐V‐AS/7721 cells. To investigate the effect of ATRA on homocysteine metabolism, cells were challenged with exogenous homocysteine. In GnT‐V‐AS/7721 cells with ATRA treatment, a significant elevation of intracellular homocysteine levels suggests that ATRA perturbs homocysteine metabolism in GnT‐V‐AS/7721 cells and, therefore, sensitizes the cells to homocysteine‐induced ER stress. An obvious increase in the levels of GRP78/Bip protein and spliced XBP1 mRNA were observed. Furthermore, we observed that ATRA blunted the homocysteine‐induced increase of GSH only in GnT‐V‐AS/7721 cells. These results demonstrate that ATRA intensifies ER stress and induces apoptosis in GnT‐V‐AS/7721 cells by disturbing homocysteine metabolism through the down‐regulation of CBS and BHMT, depleting the cellular GSH and, in turn, altering the cellular redox status. In addition, we showed that ATRA did not trigger ER stress, induce apoptosis, or affect homocysteine metabolism in L02 cells, which is a cell type that is derived from normal liver tissue. These results provide support for the hypothesis that ATRA is an anticancer agent. J. Cell. Biochem. 109: 468–477, 2010. © 2009 Wiley‐Liss, Inc.     

Mobile loop mutations in an archaeal inositol monophosphatase: Modulating three‐metal ion assisted catalysis and lithium inhibition

The inositol monophosphatase (IMPase) enzyme from the hyperthermophilic archaeon Methanocaldococcus jannaschii requires Mg²⁺ for activity and binds three to four ions tightly in the absence of ligands: K_D = 0.8 μM for one ion with a K_D of 38 μM for the other Mg²⁺ ions. However, the enzyme requires 5–10 mM Mg²⁺ for optimum catalysis, suggesting substrate alters the metal ion affinity. In crystal structures of this archaeal IMPase with products, one of the three metal ions is coordinated by only one protein contact, Asp38. The importance of this and three other acidic residues in a mobile loop that approaches the active site was probed with mutational studies. Only D38A exhibited an increased kinetic K_D for Mg²⁺; D26A, E39A, and E41A showed no significant change in the Mg²⁺ requirement for optimal activity. D38A also showed an increased K_m, but little effect on k_cat. This behavior is consistent with this side chain coordinating the third metal ion in the substrate complex, but with sufficient flexibility in the loop such that other acidic residues could position the Mg²⁺ in the active site in the absence of Asp38. While lithium ion inhibition of the archaeal IMPase is very poor (IC₅₀～250 mM), the D38A enzyme has a dramatically enhanced sensitivity to Li⁺ with an IC₅₀ of 12 mM. These results constitute additional evidence for three metal ion assisted catalysis with substrate and product binding reducing affinity of the third necessary metal ion. They also suggest a specific mode of action for lithium inhibition in the IMPase superfamily.     

Proteomic analysis on the alteration of protein expression in gills of ayu (Plecoglossus altivelis) associated with salinity change

Gill is the primary osmoregulatory organ for euryhaline fish to acclimate salinity change. The effect of salinity on gill proteome in ayu, Plecoglossus altivelis, was investigated by two-dimensional gel electrophoresis (2-DE) and matrix assisted laser desorption ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS). Eight of eighteen altered proteins were successfully identified. They are involved in osmoregulation, cytoskeleton, energy metabolism, and stress response. Our results showed that vinculin, echinoderm microtubule-associated protein like protein 1, pyruvate kinase, betaine–homocysteine methyltransferase (BHMT), transaldolase, glyceraldehyde 3-phosphate dehydrogenase, and heat shock protein 70 (HSP70) were down-regulated, whereas cofilin was up-regulated when ayu transferred from fresh water (FW) to brackish water (BW). Partial cDNA sequences of BHMT, HSP70, Na⁺/K⁺ ATPase (NKA) α-subunit and 18S rRNA genes were subsequently determined and used for 2-DE data verification by real-time PCR. Gill BHMT and HSP70 mRNAs decreased significantly in BW-transferred ayu, while NKA α-subunit mRNA had no significant change. It was suggested that cell volume-regulatory response, especially the protection by the BHMT/betaine system might play an important role in ayu acclimation to salinity change.     

Unlocking Few‐Layered Ternary Chalcogenides for High‐Performance Potassium‐Ion Storage

Potassium‐ion batteries (KIBs) have attracted increasing attention for grid‐scale energy storage due to the abundance of potassium resources, low cost, and competitive energy density. The key challenge for KIBs is to develop high‐performance electrode materials. However, the exploration of high‐capacity and ultrastable electrodes for KIBs remains challenging because of the sluggish diffusion kinetics of K⁺ ions during the charging/discharging processes. This study reports for the first time a facile ion‐intercalation‐mediated exfoliation method with Mg²⁺ cations and NO₃^– anions as ion assistants for the fabrication of expanded few‐layered ternary Ta₂NiSe₅ (EF‐TNS) flakes with interlayer spacing up to 1.1 nm and abundant Se sites (NiSe₄ tetrahedra/TaSe₆ octahedra clusters) for superior potassium‐ion storage. The EF‐TNS deliver a high capacity of 315 mAh g^–1, excellent rate capability (121 mAh g^–1 at a current density of 1000 mA g^–1), and ultrastable cycling performance (81.4% capacity retention after 1100 cycles). Detailed theoretical analysis via first‐principles calculations and experimental results elucidate that K⁺ ions intercalate through the expanded interlayers effectively and prefer to transport along zigzag pathways in layered Ta₂NiSe₅. This work provides a new avenue for designing novel ternary intercalation/pseudocapacitance‐type KIBs with high capacity, excellent rate capability, and superior long‐term cycling performance.     

Betaine analogues alter homocysteine metabolism in rats

Glycine betaine supplementation lowers homocysteine levels in homocystinuria and in chronic renal failure patients through methylation catalysed by betaine-homocysteine methyltransferase (BHMT). The aim of this study was to determine the effect of glycine betaine analogues on homocysteine metabolism in Lewis rats. Glycine betaine, proline betaine, trigonelline, dimethylsulfoniopropionate (DMSP) or dimethylthetin (1.5 mmoles) was subcutaneously administered to rats fed a low betaine diet. The effect of each betaine on total plasma homocysteine and urinary and plasma betaine concentrations was monitored for 24h following administration. Baseline plasma homocysteine was 8.5 +/- micromol/l (S.E.M., n=44) and compared to controls concentrations decreased following glycine betaine (0.8+/-0.4 micromol/l, P = 0.064), DMSP (1.0+/-0.5 micromol/l, P = 0.041) and dimethylthetin (1.5 +/- 0.7micromol/l, P = 0.033) treatment, while concentrations increased following proline betaine (2.24 +/-0.7micromol/l, P = 0.002) and trigonelline (1.6 +/-0.3 micromol/l, P < 0.001) treatment. The effect of glycine betaine, DMSP and dimethylthetin on circulating homocysteine concentrations was thought to be mediated by BHMT in vivo. This hypothesis was supported by the finding that circulating glycine betaine concentrations increased following DMSP and dimethylthetin treatment. Proline betaine and trigonelline appeared to be poor BHMT substrates, being largely excreted in the urine unchanged, yet increased circulating homocysteine levels. This suggests they are inhibitors of BHMT. Urinary excretion of glycine betaine increased following treatment with all betaines, suggesting that the resorption of glycine betaine in the kidney was inhibited. The study shows that glycine betaine analogues have multiple effects on homocysteine metabolism (250).     

Luminescent properties of 4‐aminobenzo‐15‐crown‐5 after preferential binding of ferric ions in aqueous solutions

We report a combined approach that introduces the use of 4‐aminobenzo‐15‐crown‐5 (4AB15C5) for the detection of ferric(III) ions by colorimetric, ultraviolet (UV)–visible light absorption, fluorescence, and live‐cell imaging techniques along with density functional theory (DFT) calculations. We have found that 4AB15C5 is sensitive and selective for binding ferric(III) ions in aqueous solutions. DFT calculations using the polarizable continuum model have been used to explain the strong binding of the ferric ion by 4AB15C5 in aqueous solutions. The detection limit in the fluorescence quenching measurements was found to be as low as 50 μM for the ferric ion with a determined Stern–Volmer constant of 1.52 × 10⁴ M^?1. Fluorescence intensity did not change for other ions tested, Fe²⁺, Co²⁺, Mn²⁺, Mg²⁺, Zn²⁺, Ca²⁺, NH₄⁺, Na⁺, and K⁺ ions. Live‐cell fluorescence imaging was also used to check the intracellular variations in ferric ion levels. Our spectroscopic data indicated that 4AB15C5 can bind ferric ions selectively in aqueous solutions.     

Surface Amorphization of Vanadium Dioxide (B) for K‐Ion Battery

Given the merits of low cost, fast ionic transport in electrolyte, and high operating voltage, potassium ion batteries (PIBs) are promising alternatives to lithium‐ion batteries. However, developing suitable electrode materials that can reversibly accommodate large potassium ions is a great challenge. Here, guided by density functional theory (DFT) calculations, it is demonstrated that the strategy of interfacial engineering via surface amorphization of VO₂ (B) nanorods (SA‐VO₂), which results in the formation of a crystalline core/amorphous shell heterostructure, enables superior K⁺ storage performance in terms of large capacity, outstanding rate capability, and long cycle stability working as an anode for PIBs. DFT calculations reveal that the created crystalline/amorphous heterointerface in SA‐VO₂ can substantially lower the surface energy, narrow the band gap, and reduce the K⁺ diffusion barrier of VO₂ (B). These conditions enable enhanced K⁺ storage capacity and rapid K⁺/electron transfer, which result in large capacity and outstanding rate capability. Using in situ X‐ray diffraction and in situ transmission electron microscopy complemented by ex situ microscopic and spectroscopic techniques, it is unveiled that the superior cycling stability originates from the excellent phase reversibility with negligible strain response and robust mechanical behavior of SA‐VO₂ upon (de)potassiation.     

A New Strategy for High‐Voltage Cathodes for K‐Ion Batteries: Stoichiometric KVPO4F

The exploration of high‐energy‐density cathode materials is vital to the practical use of K‐ion batteries. Layered K‐metal oxides have too high a voltage slope due to their large K⁺–K⁺ interaction, resulting in low specific capacity and average voltage. In contrast, the 3D arrangement of K⁺, with polyanions separating them, reduces the strength of the effective K⁺‐K⁺ repulsion, which in turn increases specific capacity and voltage. Here, stoichiometric KVPO₄F for use as a high‐energy‐density K‐ion cathode is developed. The KVPO₄F cathode delivers a reversible capacity of ≈105 mAh g^?1 with an average voltage of ≈4.3 V (vs K/K⁺), resulting in a gravimetric energy density of ≈450 Wh kg^?1. During electrochemical cycling, the K_xVPO₄F cathode goes through various intermediate phases at x = 0.75, 0.625, and 0.5 upon K extraction and reinsertion, as determined by ex situ X‐ray diffraction characterization and ab initio calculations. This work further explains the role of oxygen substitution in KVPO_4+xF_1?x: the oxygenation of KVPO₄F leads to an anion‐disordered structure which prevents the formation of K⁺/vacancy orderings without electrochemical plateaus and hence to a smoother voltage profile.     

A Layered Inorganic–Organic Open Framework Material as a 4 V Positive Electrode with High‐Rate Performance for K‐Ion Batteries

K‐ion batteries (KIBs) are promising for large‐scale energy storage owing to various advantages like the high abundance of potassium resources in the Earth's crust, high operational potentials, and high power due to fast diffusion of K⁺ ions. However, to realize the practical application of KIBs, electrode materials are needed with high operational voltage, good capacity, long cycle life, and low‐cost. This work reports a layered open framework material, K₂[(VOHPO₄)₂(C₂O₄)], composited with reduced graphene oxide (rGO) as a 4 V positive electrode material for KIBs. The material is prepared by a simple precipitation reaction at room temperature. The material demonstrates reversible K‐extraction/insertion with conventional carbonate ester KPF₆ solutions; however, with low specific capacity and low Coulombic efficiency. A high discharge capacity of >100 mAh g^?1 with good cycling stability and higher Coulombic efficiency is achieved in a highly concentrated electrolyte, 7 mol kg^?1 of potassium bis(fluorosulfonyl)amide (KFSA) in dimethoxyethane (DME) at 0.1 C rate. Due to the facile migration of K⁺ ions in the framework, the material exhibits excellent rate capability with a discharge capacity of 80 mAh g^?1 at 10 C rate, and a good capacity retention of 67% after 500 cycles at 2 C rate.     

Nucleotide recognition by CopA,a Cu+‐transporting P‐type ATPase

Heavy metal pumps constitute a large subgroup in P‐type ion‐transporting ATPases. One of the outstanding features is that the nucleotide binding N‐domain lacks residues critical for ATP binding in other well‐studied P‐type ATPases. Instead, they possess an HP‐motif and a Gly‐rich sequence in the N‐domain, and their mutations impair ATP binding. Here, we describe 1.85 Å resolution crystal structures of the P‐ and N‐domains of CopA, an archaeal Cu⁺‐transporting ATPase, with bound nucleotides. These crystal structures show that CopA recognises the adenine ring completely differently from other P‐type ATPases. The crystal structure of the His462Gln mutant, in the HP‐motif, a disease‐causing mutation in human Cu⁺‐ATPases, shows that the Gln side chain mimics the imidazole ring, but only partially, explaining the reduction in ATPase activity. These crystal structures lead us to propose a role of the His and a mechanism for removing Mg²⁺ from ATP before phosphoryl transfer.     

Synthesis,characterization and sol–gel entrapment of a crown ether‐styryl fluoroionophore

The synthesis and initial evaluation of a new dye‐functionalized crown‐ether, 2‐[2‐(2,3,5,6,8,9,11,12,14,15‐decahydro‐1,4,7,10,13,16‐benzohexaoxacyclooctadecin)ethenyl]‐3‐methyl benzothiazolium iodide (denoted BSD), are reported. This molecule contains a benzyl 18‐crown‐6 moiety as the ionophore and a benzothiazolium to spectrally transduce ion binding. Binding of K⁺ to BSD in methanol causes shifts in the both absorbance and fluorescence emission maxima, as well as changes in the molar absorptivity and the emission intensity. Apparent dissociation constants (K_d) in the range 30–65 µ m were measured. In water and neutral buffer, K_d values were approximately 1 m m . BSD was entrapped in sol–gel films composed of methyltriethoxysilane (MTES) and tetraethylorthosilicate (TEOS) with retention of its spectral properties and minimal leaching. K⁺ binding to BSD in sol–gel films immersed in pH 7.4 buffer causes significant fluorescence quenching, with an apparent response time of approximately 2 min and an apparent K_d of 1.5 m m . Copyright © 2009 John Wiley & Sons, Ltd.     

Localized High‐Concentration Electrolytes Boost Potassium Storage in High‐Loading Graphite

Reversible intercalation of potassium‐ion (K⁺) into graphite makes it a promising anode material for rechargeable potassium‐ion batteries (PIBs). However, the current graphite anodes in PIBs often suffer from poor cyclic stability with low coulombic efficiency. A stable solid electrolyte interphase (SEI) is necessary for stabilizing the large interlayer expansion during K⁺ insertion. Herein, a localized high‐concentration electrolyte (LHCE) is designed by adding a highly fluorinated ether into the concentrated potassium bis(fluorosulfonyl)imide/dimethoxyethane, which forms a durable SEI on the graphite surface and enables highly reversible K⁺ intercalation/deintercalation without solvent cointercalation. Furthermore, this LHCE shows a high ionic conductivity (13.6 mS cm^?1) and excellent oxidation stability up to 5.3 V (vs K⁺/K), which enables compatibility with high‐voltage cathodes. The kinetics study reveals that K⁺ intercalation/deintercalation does not follow the same pathway. The potassiated graphite exhibits excellent depotassiation rate capability, while the formation of a low stage intercalation compound is the rate‐limiting step during potassiation.     

Combined computational and biochemical study reveals the importance of electrostatic interactions between the “pH sensor” and the cation binding site of the sodium/proton antiporter NhaA of Escherichia coli

Sodium proton antiporters are essential enzymes that catalyze the exchange of sodium ions for protons across biological membranes. The crystal structure of NhaA has provided a basis to explore the mechanism of ion exchange and its unique regulation by pH. Here, the mechanism of the pH activation of the antiporter is investigated through functional and computational studies of several variants with mutations in the ion‐binding site (D163, D164). The most significant difference found computationally between the wild type antiporter and the active site variants, D163E and D164N, are low pK_a values of Glu78 making them insensitive to pH. Although in the variant D163N the pK_a of Glu78 is comparable to the physiological one, this variant cannot demonstrate the long‐range electrostatic effect of Glu78 on the pH‐dependent structural reorganization of trans‐membrane helix X and, hence, is proposed to be inactive. In marked contrast, variant D164E remains sensitive to pH and can be activated by alkaline pH shift. Remarkably, as expected computationally and discovered here biochemically, D164E is viable and active in Na⁺/H⁺ exchange albeit with increased apparent K_M. Our results unravel the unique electrostatic network of NhaA that connect the coupled clusters of the “pH sensor” with the binding site, which is crucial for pH activation of NhaA. Proteins 2009. © 2009 Wiley‐Liss, Inc.     

Realizing Reversible Conversion‐Alloying of Sb(V) in Polyantimonic Acid for Fast and Durable Lithium‐ and Potassium‐Ion Storage

Finding suitable electrode materials for alkali‐metal‐ion storage is vital to the next‐generation energy‐storage technologies. Polyantimonic acid (PAA, H₂Sb₂O₆ · nH₂O), having pentavalent antimony species and an interconnected tunnel‐like pyrochlore crystal framework, is a promising high‐capacity energy‐storage material. Fabricating electrochemically reversible PAA electrode materials for alkali‐metal‐ion storage is a challenge and has never been reported due to the extremely poor intrinsic electronic conductivity of PAA associated with the highest oxidation state Sb(V). Combining nanostructure engineering with a conductive‐network construction strategy, here is reported a facile one‐pot synthesis protocol for crafting uniform internal‐void‐containing PAA nano‐octahedra in a composite with nitrogen‐doped reduced graphene oxide nanosheets (PAA?N‐RGO), and for the first time, realizing the reversible storage of both Li⁺ and K⁺ ions in PAA?N‐RGO. Such an architecture, as validated by theoretical calculations and ex/in situ experiments, not only fully takes advantage of the large‐sized tunnel transport pathways (0.37 nm²) of PAA for fast solid‐phase ionic diffusion but also leads to exponentially increased electrical conductivity (3.3 S cm^?1 in PAA?N‐RGO vs 4.8 × 10^?10 S cm^?1 in bare‐PAA) and yields an inside‐out buffer function for accommodating volume expansion. Compared to electrochemically irreversible bare‐PAA, PAA?N‐RGO manifests reversible conversion‐alloying of Sb(V) toward fast and durable Li⁺‐ and K⁺‐ion storage.     

Betaine:homocysteine methyltransferase from rat liver: purification and inhibition by a boronic acid substrate analog.

Betaine:homocysteine methyltransferase (BHMT) from rat liver has been highly purified by an efficient procedure requiring only two chromatographic steps: Sephadex G-100 chromatography and fast protein liquid chromatography chromatofocusing. A 170-fold purification and 7.5% overall yield were achieved. Chromatofocusing yielded three active forms of BHMT with pI values near 8.0, 7.6, and 7.0. The subunit molecular weight of each active form is 45,000 Da as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the native enzyme has a molecular weight of 270,000 as determined by exclusion chromatography. The stability of the purified enzyme was found to be potentiated by the presence of 1 mM dimethylglycine and 1 mM homocysteine. Boronate analogs of betaine (pinanyl N,N,N-trimethylaminomethaneboronate) (4) and dimethylglycine (pinanyl N,N-dimethylaminomethaneboronate) were synthesized from pinanyl iodomethaneboronate (3) and trimethylamine or dimethylamine, respectively. The free acid of the betaine analog (5) was reversibly generated from (4). The inhibition of BHMT by (5) appears competitive with a Ki = 45 microM. Since the Km for betaine measured with the purified enzyme is near 0.1 mM, the boronic acid analog of betaine appears to function effectively as a substrate analog inhibitor of BHMT. The analog does not appear to act as a methyl donor to homocysteine when (5) is substituted for betaine in the enzyme reaction. In addition, an enzyme assay based upon C3-cyano reverse phase HPLC detection of the o-phthalaldehyde derivative of methionine was developed as an alternative to the standard radiochemical assay. Betaine:homocysteine methyltransferase in the picomole range can be quantitated using this assay as indicated by a linear response of enzyme activity to protein concentration.     

Intrinsically disordered inhibitor of glutamine synthetase is a functional protein with random‐coil‐like pKa values

The sequential action of glutamine synthetase (GS) and glutamate synthase (GOGAT) in cyanobacteria allows the incorporation of ammonium into carbon skeletons. In the cyanobacterium Synechocystis sp. PCC 6803, the activity of GS is modulated by the interaction with proteins, which include a 65‐residue‐long intrinsically disordered protein (IDP), the inactivating factor IF7. This interaction is regulated by the presence of charged residues in both IF7 and GS. To understand how charged amino acids can affect the binding of an IDP with its target and to provide clues on electrostatic interactions in disordered states of proteins, we measured the pK_a values of all IF7 acidic groups (Glu32, Glu36, Glu38, Asp40, Asp58, and Ser65, the backbone C‐terminus) at 100 mM NaCl concentration, by using NMR spectroscopy. We also obtained solution structures of IF7 through molecular dynamics simulation, validated them on the basis of previous experiments, and used them to obtain theoretical estimates of the pK_a values. Titration values for the two Asp and three Glu residues of IF7 were similar to those reported for random‐coil models, suggesting the lack of electrostatic interactions around these residues. Furthermore, our results suggest the presence of helical structure at the N‐terminus of the protein and of conformational changes at acidic pH values. The overall experimental and in silico findings suggest that local interactions and conformational equilibria do not play a role in determining the electrostatic features of the acidic residues of IF7.     

High homocysteine induces betaine depletion

Betaine is the substrate of the liver- and kidney-specific betaine-homocysteine (Hcy) methyltransferase (BHMT), an alternate pathway for Hcy remethylation. We hypothesized that BHMT is a major pathway for homocysteine removal in cases of hyperhomocysteinaemia (HHcy). Therefore, we measured betaine in plasma and tissues from patients and animal models of HHcy of genetic and acquired cause. Plasma was collected from patients presenting HHcy without any Hcy interfering treatment. Plasma and tissues were collected from rat models of HHcy induced by diet and from a mouse model of cystathionine β-synthase (CBS) deficiency. S-adenosyl-methionine (AdoMet), S-adenosyl-homocysteine (AdoHcy), methionine, betaine and dimethylglycine (DMG) were quantified by ESI—LC–MS/MS. mRNA expression was quantified using quantitative real-time (QRT)-PCR. For all patients with diverse causes of HHcy, plasma betaine concentrations were below the normal values of our laboratory. In the diet-induced HHcy rat model, betaine was decreased in all tissues analysed (liver, brain, heart). In the mouse CBS deficiency model, betaine was decreased in plasma, liver, heart and brain, but was conserved in kidney. Surprisingly, BHMT expression and activity was decreased in liver. However, in kidney, BHMT and SLC6A12 expression was increased in CBS-deficient mice. Chronic HHcy, irrespective of its cause, induces betaine depletion in plasma and tissues (liver, brain and heart), indicating a global decrease in the body betaine pool. In kidney, betaine concentrations were not affected, possibly due to overexpression of the betaine transporter SLC6A12 where betaine may be conserved because of its crucial role as an osmolyte.     

Potential anti‐bacterial drug target: Structural characterization of 3,4‐dihydroxy‐2‐butanone‐4‐phosphate synthase from Salmonella typhimurium LT2

3,4‐Dihydroxy‐2‐butanone‐4‐phosphate synthase (DHBPS) encoded by ribB gene is one of the first enzymes in riboflavin biosynthesis pathway and catalyzes the conversion of ribulose‐5‐phosphate (Ru5P) to 3,4‐dihydroxy‐2‐butanone‐4‐phosphate and formate. DHBPS is an attractive target for developing anti‐bacterial drugs as this enzyme is essential for pathogens, but absent in humans. The recombinant DHBPS enzyme of Salmonella requires magnesium ion for its activity and catalyzes the formation of 3,4‐dihydroxy‐2‐butanone‐4‐phosphate from Ru5P at a rate of 199 nmol min^?1 mg^?1 with K_m value of 116 μM at 37°C. Further, we have determined the crystal structures of Salmonella DHBPS in complex with sulfate, Ru5P and sulfate‐zinc ion at a resolution of 2.80, 2.52, and 1.86 Å, respectively. Analysis of these crystal structures reveals that the acidic loop (residues 34–39) responsible for the acid‐base catalysis is disordered in the absence of substrate or metal ion at the active site. Upon binding either substrate or sulfate and metal ions, the acidic loop becomes stabilized, adopts a closed conformation and interacts with the substrate. Our structure for the first time reveals that binding of substrate Ru5P alone is sufficient for the stabilization of the acidic active site loop into a closed conformation. In addition, the Glu38 residue from the acidic active site loop undergoes a conformational change upon Ru5P binding, which helps in positioning the second metal ion that stabilizes the Ru5P and the reaction intermediates. This is the first structural report of DHBPS in complex with either substrate or metal ion from any eubacteria. Proteins 2010. © 2010 Wiley‐Liss, Inc.     

Nanosheets‐Assembled CuSe Crystal Pillar as a Stable and High‐Power Anode for Sodium‐Ion and Potassium‐Ion Batteries

Thanks to low costs and the abundance of the resources, sodium‐ion (SIBs) and potassium‐ion batteries (PIBs) have emerged as leading candidates for next‐generation energy storage devices. So far, only few materials can serve as the host for both Na⁺ and K⁺ ions. Herein, a cubic phase CuSe with crystal‐pillar‐like morphology (CPL‐CuSe) assembled by the nanosheets are synthesized and its dual functionality in SIBs and PIBs is comprehensively studied. The electrochemical measurements demonstrate that CPL‐CuSe enables fast Na⁺ and K⁺ storage as well as the sufficiently long duration. Specifically, the anode delivers a specific capacity of 295 mA h g^?1 at current density of 10 A g^?1 in SIBs, while 280 mA h g^?1 at 5 A g^?1 in PIBs, as well as the high capacity retention of nearly 100% over 1200 cycles and 340 cycles, respectively. Remarkably, CPL‐CuSe exhibits a high initial coulombic efficiency of 91.0% (SIBs) and 92.4% (PIBs), superior to most existing selenide anodes. A combination of in situ X‐ray diffraction and ex situ transmission electron microscopy tests fundamentally reveal the structural transition and phase evolution of CuSe, which shows a reversible conversion reaction for both cells, while the intermediate products are different due to the sluggish K⁺ insertion reaction.     

The Potassium Requirement for Growth and Embryogenesis in Wild Carrot Suspension Cultures

A requirement for potassium for growth and forembryogenesis in suspension cultures of wild carrot (Daucus carota L.) was demonstrated. The concentration of K⁺ required for maximal growth (1 mM) was less than that required for maximal embryogenesis (20 mM). Neither Na⁺ nor NH₄⁺ could replace K⁺. Ammonium ion enhanced embryogenesis when K⁺ was present at suboptimal levels greater than 1 mM. Nitrogen sources strongly influenced growth and embryogenesis, but the effects of nitrogen were separable from those of K⁺. Subline differences were noted. Subline CSC-29 produced nearly half the maximum embryo number in 1 mM K⁺ while CSC-31 produced no embryos at that K⁺ concentration. Growth of CSC-29 was slightly repressed by Na⁺, but no more than by similarconcentrations of K⁺. Growth of CSC-31 in 1 mM K⁺ was strongly repressed by Na⁺. Embryogenesis in CSC-29 was unaffected by Na⁺. In CSC-31, Na⁺ repressed embryogenesis at lower concentrations of K⁺.