Isolation of 2′-Hydroxy-4′-methoxy propiophenone from Tribolium castaneum and T confusum

The copper binding properties were influenced by growth phase of cells, pH and concentration of copper in reaction mixtures. The efficiency of copper absorption increased with growth time and was largest at the mid-logarithmic growth phase. The time course of copper absorption was biphasic, that copper rapidly bound to cell surface for initial few minutes after addition of copper and then the copper was slowly transported into cells. The copper binding to the cell surface depended on the molecular form of copper complex in the reaction mixture and the ligand residue to copper on the cell surface. Double reciprocal plots of absorption velocity of copper vs. copper concentration gave straight lines at low concentration between 0.01 to 0.1 mm. The apparent affinity of copper to the cells of stationary growth phase was the same as that of logarithmic growth phase, that is, the Km values were about 0.01 mm. On the other hand, at high concentration of copper between 0.1 to 5.0 mm the apparent affinity decreased but the absorption velocity of copper remarkably increased. Zinc sulfate most strongly inhibited the copper absorption in this test. It was assumed that zinc competitively bound to the copper binding sites of cell surface.     

Response of Gram-positive bacteria to copper stress

The Gram-positive bacteria Enterococcus hirae, Lactococcus lactis, and Bacillus subtilis have received wide attention in the study of copper homeostasis. Consequently, copper extrusion by ATPases, gene regulation by copper, and intracellular copper chaperoning are understood in some detail. This has provided profound insight into basic principles of how organisms handle copper. It also emerged that many bacterial species may not require copper for life, making copper homeostatic systems pure defense mechanisms. Structural work on copper homeostatic proteins has given insight into copper coordination and bonding and has started to give molecular insight into copper handling in biological systems. Finally, recent biochemical work has shed new light on the mechanism of copper toxicity, which may not primarily be mediated by reactive oxygen radicals.     

Potential Phytoextraction and Phytostabilization of Perennial Peanut on Copper-Contaminated Vineyard Soils and Copper Mining Waste

This study sought to evaluate the potential of perennial peanut (Arachis pintoi) for copper phytoremediation in vineyard soils (Inceptisol and Mollisol) contaminated with copper and copper mining waste. Our results showed high phytomass production of perennial peanut in both vineyard soils. Macronutrient uptakes were not negatively affected by perennial peanut cultivated in all contaminated soils. Plants cultivated in Mollisol showed high copper concentrations in the roots and shoots of 475 and 52 mg kg⁻¹, respectively. Perennial peanut plants showed low translocation factor values for Cu, although these plants showed high bioaccumulation factor (BCF) for both vineyard soils, Inceptisol and Mollisol, with BCF values of 3.83 and 3.24, respectively, being characterized as a copper hyperaccumulator plant in these soils. Copper phytoextraction from Inceptisol soil was the highest for both roots and entire plant biomass, with more than 800 mg kg⁻¹ of copper in whole plant. The highest potential copper phytoextraction by perennial peanut was in Inceptisol soil with copper removal of 2,500 g ha⁻¹. Also, perennial peanut showed high potential for copper phytoremoval in copper mining waste and Mollisol with 1,700 and 1,500 g of copper per hectare, respectively. In addition, perennial peanuts characterized high potential for phytoextraction and phytostabilization of copper in vineyard soils and copper mining waste.     

Copper Supply in Relation to Content and Redistribution of Copper Among Organs of the Wheat Plant

The relationship of copper supply to the content and movementof copper among organs of wheat plants was examined at sevenstages in their growth from seedlings to maturity on a copperdeficient sand. In the absence of copper (Cu₀), plants becameseverely copper deficient and produced no grain; developmentof tillers, leaves, stems, and inflorescences was delayed andgrowth of roots strongly depressed; leaf senescence was retardedand tiller growth was prolonged. Application of a marginal supplyof copper (Cu₁) overcame all symptoms and promoted growth andgrain production. Increasing copper supply eightfold (Cu₂) didnot change vegetative or grain production. Copper concentrations in stems, individual leaves, and wholetops were highest and responded most strongly to copper supplywhen they were young. As they aged, Cu₁ and Cu₂ leaves lostcopper rapidly; the first Cu₀ leaves retained their copper andremained healthy for more than 7 weeks even though younger leavesdeveloped severe copper deficiency. In all treatments, lossof copper from the oldest leaf paralleled senescence and theloss of nitrogen. It is suggested that copper does not move out of plant leavesuntil they lose organic nitrogen compounds. As a result, copperbehaves in non-senescent leaves as if it is not mobile in plantphloem. But under conditions favouring senescence, copper ishighly mobile: in the present experiment, 67 per cent of thecopper present in vegetative organs of the Cu₂ primary shootat flowering moved from them during grain development and thiscould account for all of the copper found in the grain at maturity. The retention of copper by leaves before senescence, its rapidloss during senescence, and the effect of copper deficiencyin delaying senescence resulted in the oldest leaf of severelydeficient Cu₀ plants in the present experiment having a highercopper concentration than that of copper adequate Cu₁ and Cu₂plants. This behaviour could account for the many reports ofanomalous C-shaped ‘Piper-Steenbjerg’ curves inthe relationship of yield to copper concentrations in planttops. The coupling of copper movement from leaves to nitrogenmovement can also account for the unusually high values reportedfor critical concentrations of copper in tops of plants givenhigh levels of nitrogen fertilizers. Old organs should not be included in samples for diagnosis ofcopper deficiency. Only young organs should be used. In thepresent experiment, the copper concentration of young leavesgave a good indication of the copper status of wheat: a valueof 1 µg g^–1 in young leaves indicated copper deficiency. copper, nitrogen, phloem transport, mineral transport, deficiency diagnosis, wheat, Triticum aestivum L.     

Porins Increase Copper Susceptibility of Mycobacterium tuberculosis

Copper resistance mechanisms are crucial for many pathogenic bacteria, including Mycobacterium tuberculosis, during infection because the innate immune system utilizes copper ions to kill bacterial intruders. Despite several studies detailing responses of mycobacteria to copper, the pathways by which copper ions cross the mycobacterial cell envelope are unknown. Deletion of porin genes in Mycobacterium smegmatis leads to a severe growth defect on trace copper medium but simultaneously increases tolerance for copper at elevated concentrations, indicating that porins mediate copper uptake across the outer membrane. Heterologous expression of the mycobacterial porin gene mspA reduced growth of M. tuberculosis in the presence of 2.5 μM copper by 40% and completely suppressed growth at 15 μM copper, while wild-type M. tuberculosis reached its normal cell density at that copper concentration. Moreover, the polyamine spermine, a known inhibitor of porin activity in Gram-negative bacteria, enhanced tolerance of M. tuberculosis for copper, suggesting that copper ions utilize endogenous outer membrane channel proteins of M. tuberculosis to gain access to interior cellular compartments. In summary, these findings highlight the outer membrane as the first barrier against copper ions and the role of porins in mediating copper uptake in M. smegmatis and M. tuberculosis.     

Isopods as indicators of the copper content of soil and litter

Summary In two species of isopods (mainlyTracheoniscus rathkei, plus a fewOniscus asellus) total copper content as well as the amount of copper extractable with zinc-dibenzyldithiocarbamate in CCl₄ (CTC) were determined. Both copper fractions show near perfect relationship with total copper concentration of the litter collected in the isopods' habitats. Between copperrich and copper-poor sites in Tirol, Austria, mean total copper content of the isopods varies by a factor of 7, mean CTC-extracted copper by a factor of 140 (Table 2). With the exception of one, particularly impoverished, site the concentration of copper in the CTC-extracted compartment reflects the total copper concentration of the food of the animals. Both copper fractions increase with the weight of animals, but the proportionality factor of the increase is three times larger for total copper than for CTC-extracted copper.A simplified geological map of Tirol is given in which the relationship between copper content of soil and litter and of the isopods at selected sites is indicated.     

Whole animal copper flux assessed by positron emission tomography in the Long – Evans cinnamon rat – a feasibility study

Copper is an essential trace element. However, excess copper can lead to oxidation of biomolecules and cell damage and copper levels must be carefully controlled. While copper homeostasis has been studied extensively at the cellular level, short-term body copper fluxes are poorly understood. Here, we assessed for the first time the feasibility of measuring whole body copper flux by positron emission tomography, using ⁶⁴Cu. A comparative approach comparing the Long – Evans cinnamon (LEC) rat to the wild type was chosen. LEC rats are an accepted model for Wilson disease, an inherited disorder of copper excretion in humans. In LEC rats as well as in Wilson patients, the copper transporting ATPase, ATP7B, is defective. This ATPase is primarily expressed in the liver and serves in copper secretion via the bile. Dysfunction of ATP7B leads to accumulation of copper in the liver. A control and an LEC rat were transgastrically injected with 10 μg of ⁶⁴Cu and the copper flux followed for three hours by whole animal PET and concomitant collection of bile, as well as the analysis of tissue following tomography. As seen by PET, the administered copper was largely trapped in the stomach and the proximal intestine, and without a significant difference between control and LEC rat. Due to an insufficient dynamic range of the PET technology, copper which was systemically absorbed and primarily transported to the liver could only be followed by sampling and by β-counting. Biliary copper excretion ensued after 15 min in the control rat, but was absent in the LEC rat. Biliary excretion reached saturation one hour after copper administration. The trapping of orally administered copper in the gastrointestinal tract may be an important mechanism to prevent copper toxicity under conditions of a sudden, excessive copper load, which cannot be alleviated by increased biliary secretion. This trapping does however limit the utility of PET to measure whole animal copper flux. Published online December 2004     

Copper biosorption by Pseudomonas cepacia and other strains

Biosorption of copper by Pseudomonas cepacia was found to be dependent on added copper concentration. Copper uptake by the cells was rapid over the range of copper concentrations tested and complete within the first 10 min of incubation time. The effect of pH on copper uptake by P. cepacia was determined using overlapping buffers over the pH range 3–8, and copper biosorption from a 10 mM copper solution was greatest at pH 7. Copper uptake (measured by analysis of cell digests) was unaffected by cyanide and azide (up to 30 mM) and by incubation of cells with a 10 mM copper solution at 4 °C. Evidence from these results suggested that copper uptake by P. cepacia cells involves surface binding and not intracellular accumulation by active transport. Biosorption of copper by various Pseudomonas isolates from metal-contaminated environments agreed well with copper biosorption by Pseudomonas strains from the National Collection of Type Cultures (NCTC).     

Copper sulfide precipitation by yeasts from Acid mine-waters

Two strains of Rhodotorula and one of Trichosporon precipitated dissolved copper with H₂S formed by reducing elemental sulfur with glucose. Iron stimulated this activity under certain conditions. In the case of Rhodotorula strain L, iron stimulated copper precipitation aerobically at a copper concentration of 18 but not 180 μg/ml. Anaerobically, the L strain required iron for precipitation of copper from a medium with 180 μg of copper per ml. Rhodotorula strain L was able to precipitate about five times as much copper anaerobically as aerobically. The precipitated copper was identified as copper sulfide, but its exact composition could not be ascertained. Iron was not precipitated by the H₂S formed by any of the yeasts. Added as ferric iron, it was able to redissolve copper sulfide formed aerobically by Rhodotorula strain L from 18 but not 180 μg of copper per ml of medium. Since the yeasts were derived from acid mine-waters, their ability to precipitate copper may be of geomicrobial importance.     

“Pulling the plug” on cellular copper: The role of mitochondria in copper export

Mitochondria contain two enzymes, Cu,Zn superoxide dismutase (Sod1) and cytochrome c oxidase (CcO), that require copper as a cofactor for their biological activity. The copper used for their metallation originates from a conserved, bioactive pool contained within the mitochondrial matrix, the size of which changes in response to either genetic or pharmacological manipulation of cellular copper status. Its dynamic nature implies molecular mechanisms exist that functionally couple mitochondrial copper handling with other, extramitochondrial copper trafficking pathways. The recent finding that mitochondrial proteins with established roles in CcO assembly can also effect changes in cellular copper levels by modulating copper efflux from the cell supports a mechanistic link between organellar and cellular copper metabolism. However, the proteins and molecular mechanisms that link trafficking of copper to and from the organelle with other cellular copper trafficking pathways are unknown. This review documents our current understanding of copper trafficking to, and within, the mitochondrion for metallation of CcO and Sod1; the pathways by which the two copper centers in CcO are formed; and, the interconnections between mitochondrial function and the regulation of cellular copper homeostasis.     

Heterozygous <Emphasis Type="BoldItalic">tx</Emphasis> mice have an increased sensitivity to copper loading: Implications for Wilson’s disease carriers

Wilson’s disease carriers constitute 1% of the human population. It is unknown whether Wilson’s disease carriers are at increased susceptibility to copper overload when exposed to chronically high levels of ingested copper. This study investigated the effect of chronic excess copper in drinking water on the heterozygous form of the Wilson's disease mouse model – the toxic milk (tx) mouse. Mice were provided with drinking water containing 300 mg/l copper for 4–7, 8–11, 12–15 or 16–20 months. At the completion of the study liver, spleen, kidney and brain tissue were analyzed by atomic absorption spectroscopy to determine copper concentration. Plasma ceruloplasmin oxidase activity and liver histology were also assessed. Chronic copper loading resulted in significantly increased liver copper in both tx heterozygous and tx homozygous mice, while wild type mice were resistant to the effects of copper loading. Copper loading effects were greatest in tx homozygous mice, with increased extrahepatic copper deposition in spleen and kidney – an effect absent in heterozygote and wild type mice. Although liver histology in homozygous mice was markedly abnormal, no histological differences were noted between heterozygous and wild type mice with copper loading. Tx heterozygous mice have a reduced ability to excrete excess copper, indicating that half of the normal liver Atp7b copper transporter activity is insufficient to deal with large copper intakes. Our results suggest that Wilson’s disease carriers in the human population may be at increased risk of copper loading if chronically exposed to elevated copper in food or drinking water.     

The Enterococcus hirae paradigm of copper homeostasis: Copper chaperone turnover, interactions, and transactions

The cop operon is a key element of copper homeostasis in Enterococcus hirae. It encodes two copper ATPases, CopA and CopB, the CopY repressor, and the CopZ metallochaperone. The cop operon is induced by copper, which allows uncompromised growth in up to 5 mM ambient copper. Copper uptake appears to be accomplished by the CopA ATPase, a member of the heavy metal CPx-type ATPases and closely related to the human Menkes and Wilson ATPases. The related CopB ATPase extrudes copper when it reaches toxic levels. Intracellular copper routing is accomplished by the CopZ copper chaperone. Using surface plasmon resonance analysis, it was demonstrated that CopZ interacts with the CopA ATPase where it probably becomes copper loaded. CopZ in turn can donate copper to the copper responsive repressor CopY, thereby releasing it from DNA. In high copper, CopZ is proteolyzed. Cell extracts were found to contain a copper activated proteolytic activity that degrades CopZ in vitro. This post-translational control of CopZ expression presumably serves to avoid the accumulation of detrimental Cu-CopZ levels.     

Metallic copper corrosion rates, moisture content, and growth medium influence survival of copper ion-resistant bacteria

The rapid killing of various bacteria in contact with metallic copper is thought to be influenced by the influx of copper ions into the cells, but the exact mechanism is not fully understood. This study showed that the kinetics of contact killing of copper surfaces depended greatly on the amount of moisture present, copper content of alloys, type of medium used, and type of bacteria. We examined antibiotic- and copper ion-resistant strains of Escherichia coli and Enterococcus faecium isolated from pig farms following the use of copper sulfate as feed supplement. The results showed rapid killing of both copper ion-resistant E. coli and E. faecium strains when samples in rich medium were spread in a thin, moist layer on copper alloys with 85% or greater copper content. E. coli strains were rapidly killed under dry conditions, while E. faecium strains were less affected. Electroplated copper surface corrosion rates were determined from electrochemical polarization tests using the Stern–Geary method and revealed decreased corrosion rates with benzotriazole and thermal oxide coating. Copper ion-resistant E. coli and E. faecium cells suspended in 0.8% NaCl showed prolonged survival rates on electroplated copper surfaces with benzotriazole coating and thermal oxide coating compared to surfaces without anti-corrosion treatment. Control of surface corrosion affected the level of copper ion influx into bacterial cells, which contributed directly to bacterial killing.     

Copper transport and Alzheimer’s disease

This brief review discusses copper transport in humans, with an emphasis on knowledge learned from one of the simplest model organisms, yeast. There is a further focus on copper transport in Alzheimer’s Disease (AD). Copper homeostasis is essential for the well-being of all organisms, from bacteria to yeast to humans: survival depends on maintaining the required supply of copper for the many enzymes, dependent on copper for activity, while ensuring that there is no excess free copper, which would cause toxicity. A virtual orchestra of proteins are required to achieve copper homeostasis. For copper uptake, Cu(II) is first reduced to Cu(I) via a membrane-bound reductase. The reduced copper can then be internalised by a copper transporter where it is transferred to copper chaperones for transport and specific delivery to various organelles. Of significance are internal copper transporters, ATP7A and ATP7B, notable for their role in disorders of copper deficiency and toxicity, Menkes and Wilson’s disease, respectively. Metallothioneins and Cu/Zn superoxide dismutase can protect against excess copper in cells. It is clear too, increasing age, environmental and lifestyle factors impact on brain copper. Studies on AD suggest an important role for copper in the brain, with some AD therapies focusing on mobilising copper in AD brains. The transport of copper into the brain is complex and involves numerous players, including amyloid precursor protein, Aβ peptide and cholesterol.     

The dichotomy of mechanisms of copper accumulation in yeast induced by enhanced levels of copper as well as menadione in growth media

Abstract

A high copper accumulation is induced in the yeast Saccharomyces cerevisiae either by menadione at the level of 200 μM in the growth medium, or by elevated concentrations of copper. While the uptake, as well as the toxicity of copper, strongly depend on the zinc concentration in the medium, there is no influence of zinc on copper intake induced by menadione. The copper binding ligand d-penicillamine suppresses the accumulation of copper in the menadione systen, whereas it has virtually no effect in the copper system. Within the range of non-toxic concentrations, copper is predominantly taken up by an energy-dependent mechanism. In contrast, the accumulation in the menadione system is clearly energy-independent. Thus there exist at least two different mechanisms for the uptake of copper in the yeast Saccharomyces cerevisiae.     

Copper tolerance in bacteria requires the activation of multiple accessory pathways

Copper is a required micronutrient for bacteria and an essential cofactor for redox-active cuproenzymes. Yet, excess copper is extremely toxic, and is exploited as a bacteriocide in medical and biotechnological applications and also by the mammalian immune system. To evade copper toxicity, bacteria not only control intracellular copper homeostasis, but they must also repair the damage caused by excess copper. In this review, we summarize the bacterial cell-wide response to copper toxicity in Enterobacteria. Tapping into the abundant research data on two key organisms, Escherichia coli and Salmonella enterica, we show that copper resistance requires both the direct copper homeostatic response and also the indirect accessory pathways that deal with copper-induced damage. Since patterns of copper response are conserved through the Proteobacteria, we propose a cell-wide view of copper detoxification and copper tolerance that can be used to identify novel targets for copper-based antibacterial therapeutics.     

外源性铜对SD鼠脑内铜稳态与γ-氨基丁酸、谷氨酸含量影响的实验研究

目的：探讨脑内铜稳态与γ-氨基丁酸、谷氨酸含量之间的关联性,了解脑内铜代谢参与神经疾病的作用机制.方法：把实验动物分为8组：对照组,戊巴比妥组,腹腔注射CuCl2组（剂量分别为2 mg/kg、10 mg/kg、50 mg/kg）,CuCl2-戊巴比妥混合组（先腹腔注射CuCl2,再注射戊巴比妥）.检测SD鼠血清和海马内不同形态铜、γ-氨基丁酸和谷氨酸含量.结果：单一铜胁迫下发现随着外源铜升高血清和海马总铜及血清非蛋白结合铜和海马内谷氨酸含量显著升高,在50 mg/kg注射剂量下达到最高（P＜0.02）;海马非蛋白结合铜和神经递质γ-氨基丁酸含量在2 mg/kg注射剂量下达到最高（P＜0.02）,并随着外源铜浓度升高而降低.单一注射戊巴比妥后海马游离铜含量明显降低（P＜0.02）.铜胁迫1小时后注射1％戊巴比妥结果表明,戊巴比妥能明显降低铜胁迫下海马及血清总铜和游离铜含量（P＜0.05）.结论：外源铜能改变体内铜稳态;铜稳态失衡导致神经递质γ-氨基丁酸、谷氨酸含量变化,γ-氨基丁酸含量与非蛋白结合铜呈正相关.     

The Consequence of Selection for Copper Tolerance on the Uptake and Accumulation of Copper inMimulus guttatus

Previous research has shown that copper tolerance inMimulusguttatusFischer ex DC. is controlled by a single major geneand can be enhanced by a number of minor genes (or modifiers).Here we report the uptake of copper by three lines which allpossessed the major tolerance gene but differed in the modifiergenes: the major gene only (IT), the major gene plus increased(HT6) and decreased (LT6) modifiers. HT6 showed the highestcopper tolerance and IT the lowest. Copper uptake was investigatedat five copper concentrations and over 30 d to analyse the concentrationof copper accumulated in roots and shoots and the partitioningof apoplastic and symplastic root copper. Significant differenceswere found for root copper concentration with the IT line accumulatingthe highest levels. Fifty-two per cent of the root copper inthe IT line is symplastic and this increases to 60% in LT6 and64% in HT6. Significant differences were recorded for shootcopper concentration with HT6 accumulating the highest and ITthe lowest. At the highest external copper concentration theHT line accumulated nearly 800 µg g^-1in its shoot, approachinglevels reported for copper hyperaccumulation.Copyright 1997Annals of Botany Company Copper tolerance; copper uptake; copper minor genes; Mimulus guttatus