BioMetals (v.25, #2)

Iron trafficking system in Helicobacter pylori by Ruiguang Ge; Xuesong Sun (247-258).
Helicobacter pylori infections are closely associated with peptic ulcers, gastric malignancy and iron deficiency anemia. Iron is essential for almost all living organisms and the investigation of iron uptake and trafficking system is thus important to understand the pathological roles of H. pylori. Up to now, the iron trafficking system of H. pylori is not yet fully clear and merits further efforts in this regards. The available information about iron uptake and regulation has been discussed in this concise review, such as FeoB in ferrous transportation, FrpB2 in hemoglobin uptake, HugZ in heme processing, virulence factors (VacA and CagA) in transferrin utilization, Pfr and NapA in iron storage and Fur in iron regulation. The identified iron trafficking system will help us to understand the pathological roles of H. pylori in the various gastric diseases and iron deficiency anemia and stimulates further development of effective anti-bacterial drugs.
Keywords: Helicobacter pylori ; Iron; Iron deficiency anemia; Gastric disease; Fur

Ferritin iron loading was studied in the presence of physiological serum phosphate concentrations (1 mM), elevated serum concentrations (2–5 mM), and intracellular phosphate concentrations (10 mM). Experiments compared iron loading into homopolymers of H and L ferritin with horse spleen ferritin. Prior to studying the reactions with ferritin, a series of control reactions were performed to study the solution chemistry of Fe2+ and phosphate. In the absence of ferritin, phosphate catalyzed Fe2+ oxidation and formed soluble polymeric Fe(III)-phosphate complexes. The Fe(III)-phosphate complexes were characterized by electron microscopy and atomic force microscopy, which revealed spherical nanoparticles with diameters of 10–20 nm. The soluble Fe(III)-phosphate complexes also formed as competing reactions during iron loading into ferritin. Elemental analysis on ferritin samples separated from the Fe(III)-phosphate complexes showed that as the phosphate concentration increased, the iron loading into horse ferritin decreased. The composition of the mineral that does form inside horse ferritin has a higher iron/phosphate ratio (~1:1) than ferritin purified from tissue (~10:1). Phosphate significantly inhibited iron loading into L ferritin, due to the lack of the ferroxidase center in this homopolymer. Spectrophotometric assays of iron loading into H ferritin showed identical iron loading curves in the presence of phosphate, indicating that the ferroxidase center of H ferritin efficiently competes with phosphate for the binding and oxidation of Fe2+. Additional studies demonstrated that H ferritin ferroxidase activity could be used to oxidize Fe2+ and facilitate the transfer of the Fe3+ into apo transferrin in the presence of phosphate.
Keywords: Non-transferrin bound iron; Ferritin; Chronic kidney disease; Transferrin; Soluble Fe(III)-phosphate complexes; Ferroxidase activity; Ferroxidase assay

Vibrio vulnificus, the cause of septicemia and serious wound infection in humans and fishes, require iron for its pathogenesis. Hemin uptake through the outer membrane receptor, HupA, is one of its many mechanisms by which it acquires iron. We report here the identification of an additional TonB-dependent hemin receptor HvtA, that is needed in conjunction with the HupA protein for optimal hemin utilization. The HvtA protein is significantly homologous to other outer membrane hemin receptors and its expression in trans restored the uptake of hemin and hemoglobin, the latter to a weaker extent, in a mutant strain that was defective in both receptors. Quantitative RT-PCR suggested that transcription of the hvtA gene was iron regulated. The operon containing the hvtA gene is homologous to the operon in V. cholerae containing the hemin receptor gene hutR suggesting a vertical transmission of the hvtA cluster from V. cholerae to V. vulnificus.
Keywords: Iron; Hemin; Hemoglobin; Outer membrane protein

Siderophore production by actinomycetes isolates from two soil sites in Western Australia by Joanna Lee; Armin Postmaster; Hooi Peng Soon; David Keast; Kerry C. Carson (285-296).
The actinomycetes are metabolically flexible soil micro-organisms capable of producing a range of compounds of interest, including siderophores. Siderophore production by actinomycetes sampled from two distinct and separate geographical sites in Western Australia were investigated and found to be generally similar in the total percentage of siderophore producers found. The only notable difference was the proportion of isolates producing catechol siderophores with only 3% found in site 1 (from the north-west of Western Australia and reportedly containing 40% magnetite) and 17% in site 2 (a commercial stone fruit orchard in the hills east of Perth with a soil base ranging from sandy loam to laterite). Further detailed characterization of isolates of interest identified a Streptomyces that produced extracellularly excreted enterobactin, the characteristic Enterobacteriaceae siderophore, and also revealed some of the conditions required for enterobactin production. Carriage of the entF gene, which codes for the synthetase responsible for the final assembly of the tri-cyclic structure of enterobactin, was confirmed by PCR in this isolate. Another separate Streptomyces produced a compound that matched the UV/VIS spectra of heterobactin, a siderophore previously only described in Rhodococcus and Nocardia.
Keywords: Siderophores; Actinomycetes; Streptomyces ; Enterobactin; Heterobactin

Dietary selenium regulation of transcript abundance of selenoprotein N and selenoprotein W in chicken muscle tissues by Jiu-li Zhang; Jin-Long Li; Xiao-dan Huang; Sun Bo; Wang Rihua; Shu Li; Shi-wen Xu (297-307).
Selenium (Se), selenoprotein N (SelN) and selenoprotein W (SelW) play a crucial role in muscle disorders. Se status highly regulates selenoprotein mRNA levels. However, few attempts have been performed on the effect of dietary Se supplementation on muscle SelN and SelW mRNA levels in birds. To investigate the effects of Se on the regulation of SelN and SelW mRNA levels in muscle tissues, one-day-old male chickens were fed either a commercial diet or a Se-supplemented diet containing 1.0, 2.0, 3.0 or 5.0 mg/kg sodium selenite for 90 days. Muscle tissues (breast, flight, thigh, shank and cardiac muscles) were collected and examined for Se content and mRNA levels of SelN and SelW. Moreover, Selenophosphate synthetase-1 (SPS-1) and selenocysteine-synthase (SecS) mRNA levels were analyzed. Significant increases in SelN mRNA levels were obtained in breast, thigh and shank muscles treated with Se, with maximal effects at 3.0 mg Se/kg diet, but 2.0 mg Se/kg diet resulted in peak levels of Sel N mRNA in flight muscles. Changes in SelW mRNA abundance in thigh and shank muscles increased in response to Se supply. After reaching a maximal level, higher Se supplementation led to a reduction in both SelN and SelW mRNAs. However, SelN and SelW mRNA levels displayed a different expression pattern in different skeletal and cardiac muscles. Thus, it suggested that skeletal and cardiac muscles SelN and SelW mRNA levels were highly regulated by Se supplementation and different muscle tissues showed differential sensitivity. Moreover, Se supplementation also regulated the levels of SPS1 and SecS mRNAs. The mRNA levels of SPS1 and SecS were enhanced in the Se supplemented groups. These data indicate that Se regulates the expression of SelN and SelW gene and affect the mRNA levels of SecS and SPS1.
Keywords: Chicken; Selenium; Selenoprotein N; Selenoprotein W; Selenophosphate synthetase-1; Selenocysteine synthase; mRNA expression; Muscle tissues

Hexavalent chromium-induced erythrocyte membrane phospholipid asymmetry by Adrian Lupescu; Kashif Jilani; Christine Zelenak; Mohanad Zbidah; Syed M. Qadri; Florian Lang (309-318).
Hexavalent (VI) chromium is a global contaminant with cytotoxic activity. Chromium (VI) induces oxidative stress, inflammation, cell proliferation, malignant transformation and may trigger carcinogenesis and at the same time apoptosis. The toxic effects of chromium (VI) at least partially result from mitochondrial injury and DNA damage. Erythrocytes lack mitochondria and nuclei but may experience an apoptosis-like suicidal cell death, i.e. eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the cell surface. Eryptosis may result from increase of cytosolic Ca2+ activity, ATP depletion and/or ceramide formation. The present study explored, whether chromium (VI) triggers eryptosis. Fluo-3-fluorescence was employed to determine cytosolic Ca2+-concentration, forward scatter to estimate cell volume, binding of fluorescent annexin V to detect phosphatidylserine exposure, hemoglobin concentration in the supernatant to quantify hemolysis, luciferin–luciferase to determine cytosolic ATP concentration and fluorescent anti-ceramide antibodies to uncover ceramide formation. A 48 h exposure to chromium (VI) (≥10 μM) significantly increased cytosolic Ca2+-concentration, decreased ATP concentration (20 μM), decreased forward scatter, increased annexin V-binding and increased (albeit to a much smaller extent) hemolysis. Chromium (VI) did not significantly modify ceramide formation. The effect of 20 μM chromium (VI) on annexin V binding was partially reversed in the nominal absence of Ca2+. The present observations disclose a novel effect of chromium (VI), i.e. Ca2+ entry and cytosolic ATP depletion in erythrocytes, effects resulting in eryptosis with cell shrinkage and cell membrane scrambling.
Keywords: Phosphatidylserine; Chromium (VI); Calcium; Cell volume; Eryptosis

Translation of the basolateral zinc transporter ZIP5 is repressed during zinc deficiency but Zip5 mRNA remains associated with polysomes and can be rapidly translated when zinc is repleted. Herein, we examined the mechanisms regulating translation of Zip5. The 3′-untranslated region (UTR) of Zip5 mRNA is well conserved among mammals and is predicted by mFOLD to form a very stable stem-loop structure. Three algorithms predict this structure to be flanked by repeated seed sites for miR-328 and miR-193a. RNAse footprinting supports the notion that a stable stem-loop structure exists in this 3′-UTR and electrophoretic mobility shift assays detect polysomal protein(s) binding specifically to the stem-loop structure in the Zip5 3′-UTR. miR-328 and miR-193a are expressed in tissues known to regulate Zip5 mRNA translation in response to zinc availability and both are polysome-associated consistent with Zip5 mRNA localization. Transient transfection assays using native and mutant Zip5 3′-UTRs cloned 3′ to luciferase cDNA revealed that the miRNA seed sites and the stem-loop function together to augment translation of Zip5 mRNA when zinc is replete.
Keywords: miRNA; mRNA; RNA stem-loop; Translation; Zinc; Zinc transporter

Transition metal abnormalities in progressive dementias by Hiroyasu Akatsu; Akira Hori; Takayuki Yamamoto; Mari Yoshida; Maya Mimuro; Yoshio Hashizume; Ikuo Tooyama; Eric M. Yezdimer (337-350).
Abnormal distributions of transition metals inside the brain are potential diagnostic markers for several central nervous system diseases, including Alzheimer’s disease (AD), Parkinson’s disease, dementia with Lewy bodies (DLB), bipolar disorders and depression. To further explore this possibility, the total concentrations of iron, zinc, copper, manganese, aluminum, chromium and cadmium were measured in post-mortem hippocampus and amygdala tissues taken from AD, DLB and Control patients. A statistically significant near fifty percent reduction in the total copper levels of AD patients was observed in both the hippocampus and amygdala. The statistical power of the hippocampus and amygdala copper analysis was found to be 86 and 74% respectively. No statistically significant deviations in the total metal concentrations were found for zinc, manganese, chromium or aluminum. Iron was found to be increased by 38% in AD amygdala tissues, but was unchanged in AD hippocampus tissues. Accounting for differences in tissue water content, as a function of both tissue type and disease state, revealed more consistencies with previous literature. To aid in the design of future experiments, the effect sizes for all tissue types and metals studied are also presented.
Keywords: Copper; Iron; Alzheimer’s disease; Lewy body; Amygdala; Hippocampus

Biogenic synthesis of multidimensional gold nanoparticles assisted by Streptomyces hygroscopicus and its electrochemical and antibacterial properties by Sathya Sadhasivam; Parthasarathi Shanmugam; Murugan Veerapandian; Ramesh Subbiah; Kyusik Yun (351-360).
The fabrication of reliable, green chemistry processes for nanomaterial synthesis is an important aspect of nanotechnology. The biosynthesis of single-pot room-temperature reduction of aqueous chloroaurate ions by Streptomyces hygroscopicus cells has been reported to facilitate the development of an industrially viable greener methodology for the synthesis of technologically important gold nanoparticles (AuNPs). Multidimensional AuNPs are generated via the manipulation of key growth parameters, including solution pH and reaction time. The synthesized nanostructures are characterized by UV/Vis and energy dispersive X-ray analysis studies. Particle morphology is characterized by HRTEM, FE-SEM and BioAFM. Additionally, we have demonstrated the electrochemical and antibacterial properties of AuNPs via cyclic voltammetry analysis and a minimal inhibitory concentration assay. Owing to the drawbacks of chemical synthesis, a biological synthesis method has been developed to generate biocompatible, inexpensive and eco-friendly size-controlled nanoparticles.
Keywords: Gold nanoparticle; Nanostructure; Multidimensional; Electrochemical

The group IIB elements, especially Cd(II) and Hg(II), are increasingly considered as potential environmental neurotoxins. This study demonstrates that the Alzheimer’s tau fragment R2, corresponding to the second repeat of the microtubule-binding domain, can bind to Zn(II), Cd(II) and Hg(II). Isothermal titration calorimetry experiments suggest that the most likely coordination site is the thiol group of Cys291, and this is further confirmed by a control experiment using a C291A mutant peptide. Circular dichroism spectrum reveals that the coordination of group IIB cations, especially Hg(II), can induce pronounced conformational conversions in natively unfolded R2, from random coil to other ordered structures. ThS fluorescence assays and electron microscopy indicate that the group IIB cations promote heparin-induced aggregation of R2, giving relatively small R2 filaments. The efficiency in promoting aggregation, as well as inducing conformational conversion, varies strongly with the cation’s polarizability. Based on these results, a model is proposed in which the cooperative folding of R2 through cross-bridging of group IIB cations is suggested to be a key factor in promoting aggregation, in addition to the effective neutralization of coulombic charge–charge repulsion by heparin, the poly-anion inducer. Our results provide clues to understanding the potential pathogenic role of group IIB metals in the development of neurofibrillary tangles, a typical hallmark of Alzheimer’s disease.
Keywords: Aggregation; Conformation; Coordination; Group IIB metals; Tau protein

Lack of ceruloplasmin expression alters aspects of copper transport to the fetus and newborn, as determined in mice by Yu-Ling Chu; Eric N. Sauble; Anthony Cabrera; Aleeza Roth; M. Leigh Ackland; Julian F. B. Mercer; Maria C. Linder (373-382).
Copper transport and accumulation were studied in virgin and lactating C57BL/6 mice, with and without expression of ceruloplasmin (Cp), to assess the importance of Cp to these processes. One hour after i.p. injection of tracer 64Cu, liver and kidney accounted for 80% of the radioactivity, and mammary gland 1%, while in lactating Cp+/+ mice 2–4 days post partum, uptake by mammary gland was 9-fold higher and that of liver and other organs was decreased, with 64Cu rapidly appearing in milk. Parallel studies in Cp−/− mice (siblings from same colony) gave virtually identical results. However, their milk contained less 64Cu, and actual copper contents determined by furnace atomic absorption were less than half those for milk from normal dams. Liver copper concentrations of pups born to Cp−/− dams also were half those of pups from wild type dams. Copper in pup brains was unaffected; but iron concentrations were reduced. We conclude that absence of Cp, while not affecting entry of exchangeable copper from the blood into the mammary gland, does have a significant effect on the availability of this metal to the newborn through the milk and in the form of stores accumulating in gestation.
Keywords: Copper; Ceruloplasmin; Copper-64; Milk; Mammary gland; Iron

Chaperones CCS, ATOX and COXIV responses to copper supplementation in healthy adults by Magdalena Araya; Monica Andrews; Fernando Pizarro; Miguel Arredondo (383-391).
Assessment of proteins in blood and other tissues has failed to identify markers of early copper effects on health. Studies in animal models show that chaperone of SOD (CCS) respond to changes of copper status. Evidence about other copper chaperones (COXIV, ATOX) is not clear. The aim of this study was to assess by means of an in vitro challenge the mRNA relative abundance of ccs, sod1, coxIV, mtIIa and atox in peripheral mononuclear cells (PMNCs) obtained from healthy individuals, acutely and chronically supplemented with small-to-moderate amounts of copper. Healthy participants received 8 mg Cu/d (supplemented group, SG) or placebo, (placebo group, PG) for 2 months. Biochemical indicators were assessed at basal (T0) and after 2 (T2) and 60 days (T60). At these times PMNCs were obtained, challenged with 1, 5 or 20 μM Cu-histidine for 20 h and the mRNA relative abundance of the selected genes assessed by real time PCR. The results showed that at T0, intracellular copper was not different between experimental and control groups. This increased at T2 and T60 when the copper in the media increased (two-way ANOVA, P < 0.001). In PG, CCS mRNA transcripts showed no significant changes (two-way ANOVA) at T2 and T60. In SG, CCS changed by treatment, time and interaction (two-way ANOVA, all P < 0.001). SOD, ATOX and COXIV expressions changed in both PG and SG showing various patterns of response, requiring further study. MTII responded as expected. We conclude that using healthy individuals as a human model, CCS but not SOD, ATOX or COXIV responded consistently to controlled changes of copper availability in an in vitro copper challenge.
Keywords: CCS; ATOX; COXIV; Copper biomarkers; Peripheral mononuclear cells

Trace metal complexation by the triscatecholate siderophore protochelin: structure and stability by James M. Harrington; John R. Bargar; Andrzej A. Jarzecki; James G. Roberts; Leslie A. Sombers; Owen W. Duckworth (393-412).
Although siderophores are generally viewed as biological iron uptake agents, recent evidence has shown that they may play significant roles in the biogeochemical cycling and biological uptake of other metals. One such siderophore that is produced by A. vinelandii is the triscatecholate protochelin. In this study, we probe the solution chemistry of protochelin and its complexes with environmentally relevant trace metals to better understand its effect on metal uptake and cycling. Protochelin exhibits low solubility below pH 7.5 and degrades gradually in solution. Electrochemical measurements of protochelin and metal–protochelin complexes reveal a ligand half-wave potential of 200 mV. The Fe(III)Proto3− complex exhibits a salicylate shift in coordination mode at circumneutral to acidic pH. Coordination of Mn(II) by protochelin above pH 8.0 promotes gradual air oxidation of the metal center to Mn(III), which accelerates at higher pH values. The Mn(III)Proto3− complex was found to have a stability constant of log β110 = 41.6. Structural parameters derived from spectroscopic measurements and quantum mechanical calculations provide insights into the stability of the Fe(III)Proto3−, Fe(III)H3Proto, and Mn(III)Proto3− complexes. Complexation of Co(II) by protochelin results in redox cycling of Co, accompanied by accelerated degradation of the ligand at all solution pH values. These results are discussed in terms of the role of catecholate siderophores in environmental trace metal cycling and intracellular metal release.
Keywords: Siderophores; Catechol; Iron; Manganese; Cobalt; Complexation

Staphylococcus aureus and Staphylococcus epidermidis are the major cause of infections associated with implanted medical devices. Colonization on abiotic and biotic surfaces is often sustained by biofilm forming strains. Human natural defenses can interfere with this virulence factor. We investigated the effect of human apo-transferrin (apo-Tf, the iron-free form of transferrin, Tf) and holo-transferrin (holo-Tf, the iron-saturated form) on biofilm formation by CA-MRSA S. aureus USA300 type (ST8-IV) and S. epidermidis (a clinical isolate and ATCC 35984 strain). Furthermore S. aureus adhesion and invasion assays were performed in a eukaryotic cell line. A strong reduction in biofilm formation with both Tfs was obtained albeit at very different concentrations. In particular, the reduction in biofilm formation was higher with apo-Tf rather than obtained with holo-Tf. Furthermore, while S. aureus adhesion to eukaryotic cells was not appreciably affected, their invasion was highly inhibited in the presence of holo-Tf, and partially inhibited by the apo form. Our results suggest that Tfs could be used as antibacterial adjuvant therapy in infection sustained by staphylococci to strongly reduce their virulence related to adhesion and cellular invasion.
Keywords: Apotransferrin; Holotransferrin; Biofilm; Cellular invasion; Internalisation

Role of the Dickeya dadantii Dps protein by Aïda Boughammoura; Dominique Expert; Thierry Franza (423-433).
During infection, the phytopathogenic enterobacterium Dickeya dadantii has to cope with iron-limiting conditions and the production of reactive oxygen species by plant cells. A tight control of the bacterial intracellular iron content is necessary for full virulence of D. dadantii: previous studies have shown that the ferritin FtnA and the bacterioferrtin Bfr, devoted to iron storage, contribute differentially to the virulence of this species. In this work, we investigated the role of the Dps miniferritin in iron homeostasis in D. dadantii. We constructed a Dps-deficient mutant by reverse genetics. This mutant grew like the wild-type stain under iron starvation and showed no decreased iron content. However, the dps mutant displayed an increased sensitivity to hydrogen peroxide in comparison to the wild-type strain. This hydrogen peroxide susceptibility only occurs when bacteria are in the stationary phase. Unlike the bfr and the ftnA mutants, the dps mutant is not affected in its pathogenicity on host plants. The dps gene expression is induced at the stationary phase of growth. The Sigma S transcriptional factor is necessary for this control. Furthermore, dps expression is positively regulated by the oxidative stress response regulator OxyR during the exponential growth phase, after hydrogen peroxide treatment. These results indicate that the Dps miniferritin from D. dadantii has a minor role in iron homeostasis, but is important in conferring tolerance to hydrogen peroxide and for survival of cells that enter the stationary phase of growth.
Keywords: Miniferritin; Resistance to oxidative stress; Gene regulation; Pathogenicity

Association of aureolic acid antibiotic, chromomycin A3 with Cu2+ and its negative effect upon DNA binding property of the antibiotic by Shibojyoti Lahiri; Toshifumi Takao; Pukhrambam Grihanjali Devi; Saptaparni Ghosh; Ayanjeet Ghosh; Amrita Dasgupta; Dipak Dasgupta (435-450).
Here we have examined the association of an aureolic acid antibiotic, chromomycin A3 (CHR), with Cu2+. CHR forms a high affinity 2:1 (CHR:Cu2+) complex with dissociation constant of 0.08 × 10−10 M2 at 25°C, pH 8.0. The affinity of CHR for Cu2+ is higher than those for Mg2+ and Zn2+ reported earlier from our laboratory. CHR binds preferentially to Cu2+ in presence of equimolar amount of Zn2+. Complex formation between CHR and Cu2+ is an entropy driven endothermic process. Difference between calorimetric and van’t Hoff enthalpies indicate the presence of multiple equilibria, supported from biphasic nature of the kinetics of association. Circular dichroism spectroscopy show that [(CHR)2:Cu2+] complex assumes a structure different from either of the Mg2+ and Zn2+ complex reported earlier. Both [(CHR)2:Mg2+] and [(CHR)2:Zn2+] complexes are known to bind DNA. In contrast, [(CHR)2:Cu2+] complex does not interact with double helical DNA, verified by means of Isothermal Titration Calorimetry of its association with calf thymus DNA and the double stranded decamer (5′-CCGGCGCCGG-3′). In order to interact with double helical DNA, the (antibiotic)2 : metal (Mg2+ and Zn2+) complexes require a isohelical conformation. Nuclear Magnetic Resonance spectroscopy shows that the Cu2+ complex adopts a distorted octahedral structure, which cannot assume the required conformation to bind to the DNA. This report demonstrates the negative effect of a bivalent metal upon the DNA binding property of CHR, which otherwise binds to DNA in presence of metals like Mg2+and Zn2+. The results also indicate that CHR has a potential for chelation therapy in Cu2+ accumulation diseases. However cytotoxicity of the antibiotic might restrict the use.
Keywords: Chromomycin; Copper ion; Electrospray ionization mass spectrometry; Multiple equilibria; DNA binding; Distorted octahedral structure

Effect of tellurite-mediated oxidative stress on the Escherichia coli glycolytic pathway by Mauricio Valdivia-González; José M. Pérez-Donoso; Claudio C. Vásquez (451-458).
To unveil the metabolic impact of tellurite in the bacterial cell, the effect of this toxicant on the expression and activity of key enzymes of the Escherichia coli glycolytic pathway was analyzed. E. coli exposure to tellurite results in: (i) increased glucose consumption, which was paralleled by an increased expression of the glucose transporter-encoding gene ptsG, (ii) augmented phosphoglucoisomerase activity and pgi transcription, (iii) decreased activity of the enzymatic regulators phosphofructokinase and pyruvate kinase. In spite of these observations, increased intracellular pyruvate, phosphoenol pyruvate and phosphorylated sugars was observed. E. coli lacking key glycolytic enzymes was considerably more sensitive to tellurite than the parental, isogenic, wild type strain. Taken together, these results suggest that increasing the availability of key metabolites (pyruvate, phosphoenol pyruvate, NADPH), required to respond to tellurite mediated-stress, E. coli shifts the carbon flux towards the pentose phosphate pathway thus facilitating the functioning of the Entner–Doudoroff pathway and/or the glycolytic productive phase.
Keywords: Tellurite; Glycolysis; Oxidative stress; Pyruvate kinase; Phosphofructokinase

Effects of Selenoprotein W gene expression by selenium involves regulation of mRNA stability in chicken embryos neurons by Jin-Long Li; Hui-Xin Li; Shu Li; Xue-Jiao Gao; Shi-Wen Xu; Zhao-Xin Tang (459-468).
Selenium (Se) and Selenoprotein W (SelW) plays a pivotal role in the brain development, function, and degeneration and that SelW expression in the brain may be affected by Se. However, the mechanism which Se regulates the SelW gene expression in neurons remains to be unclear. To investigate the effects of the SelW gene expression and mRNA stability induced by Se, primary cultured chicken embryos neurons derived from 8-day-old chick embryo cerebral hemispheres were treated with 10−9–10−5 mol/l Se as selenite for 3, 6, 12, 24 or 48 h, respectively. The morphology and viability of Neurons was detected. The SelW mRNA expression level and mRNA half-life was examined in Se-treated neurons. The relative low concentrations of Se enhanced the neurite outgrowth, increased the SelW mRNA levels and elevated the mRNA half-life of chick embryo neurons. In contrast, the high concentrations of Se presented neurotoxic to neurons, decreased the SelW mRNA levels and reduced the mRNA half-life of neuronal cells. These results suggest that the alteration of post-transcriptional stabilization of SelW mRNA is an important mechanism of Se-induced the elevation or reduction of the SelW expression level in chick embryo neurons.
Keywords: Chicken embryos neurons; Selenoprotein W; Selenium; mRNA expression; mRNA stability

Zinc enhances adiponectin oligomerization to octadecamers but decreases the rate of disulfide bond formation by David B. Briggs; Rebecca M. Giron; Karina Schnittker; Madeline V. Hart; Chad K. Park; Andrew C. Hausrath; Tsu-Shuen Tsao (469-486).
Adiponectin, a hormone secreted from adipocytes, has been shown to protect against development of insulin resistance, ischemia–reperfusion injury, and inflammation. Adiponectin assembles into multiple oligomeric isoforms: trimers, hexamers and several higher molecular weight (HMW) species. Of these, the HMW species are selectively decreased during the onset of type 2 diabetes. Despite the critical role of HMW adiponectin in insulin responsiveness, its assembly process is poorly understood. In this report, we investigated the role of divalent cations in adiponectin assembly. Purified adiponectin 18mers, the largest HMW species, did not collapse to smaller oligomers after treatment with high concentrations of EDTA. However, treatment with EDTA or another chelator DTPA inhibited the oligomerization of 18mers from trimers in vitro. Zn2+ specifically increased the formation of 18mers when compared with Cu2+, Mg2+, and Ca2+. Distribution of adiponectin oligomers secreted from zinc chelator TPEN-treated rat adipocytes skewed toward increased proportions of hexamers and trimers. While we observed presence of zinc in adiponectin purified from calf serum, the role of zinc in disulfide bonding between oligomers was examined because the process is critical for 18mer assembly. Surprisingly, Zn2+ inhibited disulfide bond formation early in the oligomerization process. We hypothesize that initial decreases in disulfide formation rates could allow adiponectin subunits to associate before becoming locked in fully oxidized conformations incapable of further oligomerization. These data demonstrate that zinc stimulates oligomerization of HMW adiponectin and possibly other disulfide-dependent protein assembly processes.
Keywords: Adiponectin; Zinc; Disulfide bonds; Adiponectin oligomerization; Macromolecular protein assembly; Adipocyte