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1.
Chemosphere ; 262: 127810, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32763578

RESUMO

Copper (Cu) is an essential element for humans and plants when present in lesser amount, while in excessive amounts it exerts detrimental effects. There subsists a narrow difference amid the indispensable, positive and detrimental concentration of Cu in living system, which substantially alters with Cu speciation, and form of living organisms. Consequently, it is vital to monitor its bioavailability, speciation, exposure levels and routes in the living organisms. The ingestion of Cu-laced food crops is the key source of this heavy metal toxicity in humans. Hence, it is necessary to appraise the biogeochemical behaviour of Cu in soil-plant system with esteem to their quantity and speciation. On the basis of existing research, this appraisal traces a probable connexion midst: Cu levels, sources, chemistry, speciation and bioavailability in the soil. Besides, the functions of protein transporters in soil-plant Cu transport, and the detrimental effect of Cu on morphological, physiological and nutrient uptake in plants has also been discussed in the current manuscript. Mechanisms related to detoxification strategies like antioxidative response and generation of glutathione and phytochelatins to combat Cu-induced toxicity in plants is discussed as well. We also delimits the Cu accretion in food crops and allied health perils from soils encompassing less or high Cu quantity. Finally, an overview of various techniques involved in the reclamation and restoration of Cu-contaminated soils has been provided.


Assuntos
Cobre/metabolismo , Produtos Agrícolas/fisiologia , Poluentes do Solo/metabolismo , Disponibilidade Biológica , Cobre/análise , Cobre/toxicidade , Produtos Agrícolas/metabolismo , Intoxicação por Metais Pesados , Humanos , Solo/química , Poluentes do Solo/análise , Poluentes do Solo/toxicidade
2.
Chemosphere ; 261: 128051, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33113650

RESUMO

The potential toxicity of copper nanoparticles (CuNPs) to early stages of fishes is not fully understood, and little is known about their effects on ionocytes and associated functions. This study used zebrafish embryos as a model to investigate the toxic effects of CuNPs on two subtypes of ionocytes. Zebrafish embryos were exposed to 0.1, 1, and 3 mg L-1 CuNPs for 96 h. After exposure, whole-body Na+ and Ca2+ contents were significantly reduced at ≥0.1 mg L-1, while the K+ content had decreased at ≥1 mg L-1. H+ and NH4+ excretion by the skin significantly decreased at ≥1 mg L-1. The number of living ionocytes labeled with rhodamine-123 had significantly decreased with ≥0.1 mg L-1 CuNPs. The ionocyte subtypes of H+-ATPase-rich (HR) and Na+/K+-ATPase-rich (NaR) cells were labeled by immunostaining and had decreased with ≥1 mg L-1. Shrinkage of the apical opening of ionocytes was revealed by scanning electronic microscopy. Functional impairment was also reflected by changes in gene expressions, including ion transporters/channels and Ca2+-regulatory hormones. This study shows that CuNP exposure can impair two subtypes of ionocytes and their associated functions, including Na+/Ca2+ uptake and H+/NH4+ excretion in zebrafish embryos.


Assuntos
Amônia/metabolismo , Cobre/toxicidade , Embrião não Mamífero/efeitos dos fármacos , Nanopartículas/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/metabolismo , Ácidos/metabolismo , Animais , Transporte Biológico , Cálcio/metabolismo , Cobre/metabolismo , Embrião não Mamífero/metabolismo , Canais Iônicos/metabolismo , Íons/metabolismo , Nanopartículas/metabolismo , Pele/metabolismo , Sódio/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Poluentes Químicos da Água/metabolismo , Proteínas de Peixe-Zebra/metabolismo
3.
Orv Hetil ; 161(35): 1488-1496, 2020 08.
Artigo em Húngaro | MEDLINE | ID: mdl-32822328

RESUMO

Copper is an essential micronutrient for the human body, taking part in several metabolic processes. It is required for the terminal oxidation, the elimination of free radicals, the proper functioning of iron metabolism, the synthesis of hormones, neurotransmitters and the stabilization of the extracellular matrix. Copper is a two-faced element. Its significance in protein-bounded form is undeniable, but free copper causes severe cell damage mainly through free radical reactions, substituting other essential metals in protein structure, and it has various effects on cell signaling pathways. The amount of copper present in the human body is part of a delicate balance. Both its deficiency and excess can develop severe symptoms and diseases. The nutritional copper balance is rarely broken, because the average daily consumption excessively covers the needs, and the body's copper stores can tolerate the temporary differences. Genetic diseases (Menkes and Wilson disease) helped to understand the copper metabolism and understand the clinical symptoms of deficiency and excessive intake. The role of copper in the development of chronic diseases is receiving increasing attention. Recent studies show its significance in neurodegenerative and cancerous diseases, both in pathogenesis and as a therapeutic target. Orv Hetil. 2020; 161(35): 1488-1496.


Assuntos
Cobre/química , Cobre/metabolismo , Radicais Livres , Degeneração Hepatolenticular/genética , Humanos , Metais , Neoplasias/genética , Oligoelementos
4.
Ecotoxicol Environ Saf ; 205: 111089, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32810645

RESUMO

Early molecular events after the exposure of heavy metals, such as aberrant DNA methylation, suggest that DNA methylation was important in regulating physiological processes for animals and accordingly could be used as environmental biomarkers. In the present study, we found that copper (Cu) exposure increased lipid content and induced the DNA hypermethylation at the whole genome level. Especially, Cu induced hypermethylation of glucose-regulated protein 78 (grp78) and peroxisome proliferator-activated receptor gamma coactivator-1α (pgc1α). CCAAT/enhancer binding protein α (C/EBPα) could bind to the methylated sequence of grp78, whereas C/EBPß could not bind to the methylated sequence of grp78. These synergistically influenced grp78 expression and increased lipogenesis. In contrast, DNA methylation of PGC1α blocked the specific protein 1 (SP1) binding and interfered mitochondrial function. Moreover, Cu increased reactive oxygen species (ROS) production, activated endoplasmic reticulum (ER) stress and damaged mitochondrial function, and accordingly increased lipid deposition. Notably, we found a new toxicological mechanism for Cu-induced lipid deposition at DNA methylation level. The measurement of DNA methylation facilitated the use of these epigenetic biomarkers for the evaluation of environmental risk.


Assuntos
Carpas/fisiologia , Cobre/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Carpas/metabolismo , Cobre/metabolismo , Estresse do Retículo Endoplasmático , Glucose/metabolismo , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Lipídeos , Metilação , Mitocôndrias/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/genética , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Ativação Transcricional , Regulação para Cima
5.
Ecotoxicol Environ Saf ; 205: 111163, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32836159

RESUMO

Arthrobacter sp. JQ-1 can completely degrade 500 mg/L of DEHP within 3 days. The minimum inhibitory concentrations (MICs) of Cu2+ could reach 1.56 mM, however, 5.0 mg/L Cu2+ apparently inhibited DEHP degradation and bacterial growth. Consequently, JQ-1 was exposed to the DEHP-copper environment to verify the toxicity mechanism based on the physiological responses of cellular multiple interfaces (cellular surface, membrane and intracellular characteristics). The results showed the combination of 500 mg/L DEHP and 5.0 mg/L Cu2+ significantly decreased cell surface hydrophobicity (CSH) and the absolute value of zeta potential, which implied the bioavailability of DEHP was decreased. The cellular surface changes were mainly due to the interaction between Cu2+ and some functional groups (CH2, CH3, aromatic rings, and amide). The weakened proton-motive force (PMF) across the plasma membrane may interfere the formation and utilization of energy, which is not conducive to the repair process of cellular damages. In this study, Non-invasive micro-test technology (NMT) was applied to the research of combined toxicity of DEHP and heavy metal ions for the first time. DEHP-copper intensified K+ efflux and Ca2+ influx across the plasma membrane, which disturbed ion homeostasis of K+ and Ca2+ and might induce apoptosis and further inhibit DEHP degradation. The decline of intracellular esterase activity indicated that the metabolic capacity is apparently restrained. This study enhances our understanding of cellular different interface processes responding to combined pollutants.


Assuntos
Arthrobacter/efeitos dos fármacos , Cobre/toxicidade , Dietilexilftalato/toxicidade , Poluentes do Solo/toxicidade , Arthrobacter/metabolismo , Arthrobacter/ultraestrutura , Biodegradação Ambiental , Cálcio/metabolismo , Cobre/metabolismo , Dietilexilftalato/metabolismo , Sinergismo Farmacológico , Potássio/metabolismo , Solo/química , Microbiologia do Solo , Poluentes do Solo/metabolismo
6.
Met Ions Life Sci ; 202020 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-32851825

RESUMO

CuA is a binuclear copper center acting as an electron transfer hub in terminal oxidases such as cytochrome c oxidase and nitrous oxide reductase. Its unique electronic structure is intimately linked to its function and has puzzled the community of biological inorganic chemistry for decades. Here we review the insights provided by different spectroscopic techniques of CuA centers, and the different experimental approaches to tackle its study, that encompass the synthesis of model compounds as well as protein engineering efforts. The contribution of the electronic structure to the thermodynamic and kinetic of electron transfer is extensively discussed. We also describe the proposed mechanism of CuAassembly in different organisms. The recent discovery of a novel CuA site opens new perspectives to this field.


Assuntos
Cobre/química , Sequência de Aminoácidos , Cobre/metabolismo , Transporte de Elétrons , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Análise Espectral
7.
Science ; 369(6499): 59-64, 2020 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-32631887

RESUMO

Eukaryotic histone H3-H4 tetramers contain a putative copper (Cu2+) binding site at the H3-H3' dimerization interface with unknown function. The coincident emergence of eukaryotes with global oxygenation, which challenged cellular copper utilization, raised the possibility that histones may function in cellular copper homeostasis. We report that the recombinant Xenopus laevis H3-H4 tetramer is an oxidoreductase enzyme that binds Cu2+ and catalyzes its reduction to Cu1+ in vitro. Loss- and gain-of-function mutations of the putative active site residues correspondingly altered copper binding and the enzymatic activity, as well as intracellular Cu1+ abundance and copper-dependent mitochondrial respiration and Sod1 function in the yeast Saccharomyces cerevisiae The histone H3-H4 tetramer, therefore, has a role other than chromatin compaction or epigenetic regulation and generates biousable Cu1+ ions in eukaryotes.


Assuntos
Cobre/metabolismo , Histonas/química , Oxirredutases/química , Multimerização Proteica , Animais , Biocatálise , Domínio Catalítico/genética , Mutação com Ganho de Função , Histonas/genética , Histonas/metabolismo , Mitocôndrias/metabolismo , Proteínas Nucleares/metabolismo , Oxirredutases/genética , Oxirredutases/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/metabolismo , Superóxido Dismutase-1/química , Fatores de Transcrição/metabolismo , Xenopus laevis
8.
Bull Environ Contam Toxicol ; 105(2): 255-260, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32632463

RESUMO

The present study investigated the biochemical toxicity and potential detoxification mechanisms in earthworms Eisenia fetida exposed to sulfamethazine (SMZ) (7.5, 15 and 30 mg kg-1) either alone or in combination with Copper (Cu) (100 mg kg-1) in soil. The results showed that increasing concentrations of SMZ in soil activated superoxide dismutase, catalase and glutathione peroxidase isozymes, suggesting reactive oxygen species (ROS) burst in earthworms. Treatment with SMZ and Cu separately or in combination caused protein oxidation and damage, elevating the synthesis of ubiquitin, the 20S proteasome, cytochrome P450 (CYP450), and heat shock protein 70 (HSP70). Such treatments also induced the activities of proteases, endoproteinase (EP) and glutathione S-transferases (GSTs). The results suggested that the ubiquitin-20S proteasome, proteases, EP and HSP70 were involved in degradation or remediation of oxidatively damaged proteins. Elevated levels of CYP450 and GSTs also participated in the detoxification of the earthworms.


Assuntos
Cobre/toxicidade , Oligoquetos/efeitos dos fármacos , Poluentes do Solo/toxicidade , Solo/química , Sulfametazina/toxicidade , Animais , Biodegradação Ambiental , Catalase/metabolismo , China , Cobre/metabolismo , Glutationa Peroxidase/metabolismo , Oligoquetos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Poluentes do Solo/metabolismo , Sulfametazina/metabolismo , Superóxido Dismutase/metabolismo
9.
Ecotoxicol Environ Saf ; 203: 110983, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32678760

RESUMO

Chelating agents have been considered as an important phytoremediation strategy to enhance heavy metal extraction from contaminated soil. A pot experiment was conducted to explore the effects of low molecular weight organic acids (LMWOAs) on the phytoremediation efficiency of copper (Cu) by castor bean, and soil enzyme activities. Results indicated that the addition of all the three kinds of LMWOAs (citric, tartaric, oxalic acids) did not decrease the biomass of castor bean, despite the fact they reduced the concentration of chlorophyll-a in leaves compared to the control. The Cu concentrations in the roots and shoots significantly increased by 6-106% and 5-148%, respectively, in the LMWOAs treatments so that the total accumulation of Cu by whole plants in all the LMWOAs treatments increased by 21-189% in comparison with the control. The values of the translocation factor (TF) and bio-concentration factor (BCF) of Cu in castor bean also rose following the addition of LMWOAs, indicating that the LMWOAs enhanced the uptake and transportation of Cu. Moreover, the application of LMWOAs did not significantly change the soil pH but significantly increased the activity of soil enzymes (urease, catalase, and alkaline phosphatase). The addition of exogenous LMWOAs increased the available Cu significantly in the soil, thus promoted the phytoextraction efficiency of Cu by castor bean. These results will provide some new insights into the practical use of LMWOAs for the phytoremediation of heavy-metal-contaminated soil employing castor bean.


Assuntos
Bioacumulação , Semente de Rícino/metabolismo , Quelantes/química , Cobre/metabolismo , Compostos Orgânicos/química , Poluentes do Solo/metabolismo , Solo/química , Ácidos/administração & dosagem , Ácidos/química , Biodegradação Ambiental , Semente de Rícino/efeitos dos fármacos , Quelantes/administração & dosagem , Peso Molecular , Compostos Orgânicos/administração & dosagem
10.
Aquat Toxicol ; 225: 105524, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32610223

RESUMO

In the present research, the effects of exposure to a sublethal concentration of zinc (Zn) on metal and ion homeostasis, and the regulation and the localization of various Zn transporters (i.e., the Zrt-Irt Like Protein (ZIP) family of Zn transporters), were investigated in zebrafish (Danio rerio) during early development. Exposure to an elevated level of Zn [4 µM (high) vs. 0.25 µM (control)] from 0 day post-fertilization (dpf) resulted in a significant increase in the whole body content of Zn at 5 dpf. A transient decrease in the whole body calcium (Ca) level was observed in 3 dpf larvae exposed to high Zn. Similarly, whole body nickel (Ni) and copper (Cu) contents were also reduced in 3 dpf larvae exposed to high Zn. Importantly, the magnitude of reduction in whole body Ni and Cu contents following Zn exposure was markedly higher than that in Ca content, suggesting that internal Ni and Cu balance were likely more sensitive to Zn exposure in developing zebrafish. Exposure to high Zn altered the mRNA expression levels of specific zip transporters, with an increase in zip1 (at 3 dpf) and zip8 (at 5 dpf), and a decrease in zip4 (at 5 dpf). The expression levels of most zip transporters tended to decrease from 3 dpf to 5 dpf with the exception of zip4 and zip8. Results from in situ hybridization revealed that several zip transporters exhibited distinct spatial distribution (e.g., zip8 in the intestinal tract, zip14 in the pronephric tubules). Overall, our findings suggested that exposure to sublethal concentrations of Zn disrupts the homeostasis of essential metals during early development and that different ZIP transporters may play unique roles in regulating Zn homeostasis in various organs in developing zebrafish.


Assuntos
Proteínas de Transporte de Cátions/genética , Homeostase/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Proteínas de Peixe-Zebra/genética , Peixe-Zebra , Zinco/toxicidade , Animais , Cálcio/metabolismo , Cobre/metabolismo , Relação Dose-Resposta a Droga , Larva/efeitos dos fármacos , Larva/metabolismo , Nível de Efeito Adverso não Observado , Poluentes Químicos da Água/metabolismo , Peixe-Zebra/fisiologia , Zinco/metabolismo
11.
ACS Appl Mater Interfaces ; 12(31): 34723-34727, 2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32657566

RESUMO

SARS-CoV-2, the virus that causes the disease COVID-19, remains viable on solids for periods of up to 1 week, so one potential route for human infection is via exposure to an infectious dose from a solid. We have fabricated and tested a coating that is designed to reduce the longevity of SARS-CoV-2 on solids. The coating consists of cuprous oxide (Cu2O) particles bound with polyurethane. After 1 h on coated glass or stainless steel, the viral titer was reduced by about 99.9% on average compared to the uncoated sample. An advantage of a polyurethane-based coating is that polyurethane is already used to coat a large number of everyday objects. Our coating adheres well to glass and stainless steel as well as everyday items that people may fear to touch during a pandemic, such as a doorknob, a pen, and a credit card keypad button. The coating performs well in the cross-hatch durability test and remains intact and active after 13 days of being immersed in water or after exposure to multiple cycles of exposure to the virus and disinfection.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/genética , Pandemias , Pneumonia Viral/genética , Betacoronavirus/patogenicidade , Cobre/química , Cobre/metabolismo , Infecções por Coronavirus/virologia , Humanos , Pneumonia Viral/virologia , Poliuretanos/química , Poliuretanos/metabolismo , Propriedades de Superfície
12.
PLoS One ; 15(7): e0235642, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32640001

RESUMO

Aspergillus tamarii grows abundantly in naturally composting waste fibers of the textile industry and has a great potential in biomass decomposition. Amongst the key (hemi)cellulose-active enzymes in the secretomes of biomass-degrading fungi are the lytic polysaccharide monooxygenases (LPMOs). By catalyzing oxidative cleavage of glycoside bonds, LPMOs promote the activity of other lignocellulose-degrading enzymes. Here, we analyzed the catalytic potential of two of the seven AA9-type LPMOs that were detected in recently published transcriptome data for A. tamarii, namely AtAA9A and AtAA9B. Analysis of products generated from cellulose revealed that AtAA9A is a C4-oxidizing enzyme, whereas AtAA9B yielded a mixture of C1- and C4-oxidized products. AtAA9A was also active on cellopentaose and cellohexaose. Both enzymes also cleaved the ß-(1→4)-glucan backbone of tamarind xyloglucan, but with different cleavage patterns. AtAA9A cleaved the xyloglucan backbone only next to unsubstituted glucosyl units, whereas AtAA9B yielded product profiles indicating that it can cleave the xyloglucan backbone irrespective of substitutions. Building on these new results and on the expanding catalog of xyloglucan- and oligosaccharide-active AA9 LPMOs, we discuss possible structural properties that could underlie the observed functional differences. The results corroborate evidence that filamentous fungi have evolved AA9 LPMOs with distinct substrate specificities and regioselectivities, which likely have complementary functions during biomass degradation.


Assuntos
Aspergillus/metabolismo , Proteínas Fúngicas/metabolismo , Glucanos/metabolismo , Oxigenases de Função Mista/metabolismo , Xilanos/metabolismo , Sequência de Aminoácidos , Sítios de Ligação , Cromatografia Líquida de Alta Pressão , Clonagem Molecular , Cobre/química , Cobre/metabolismo , Proteínas Fúngicas/classificação , Proteínas Fúngicas/genética , Glucanos/análise , Glucanos/química , Oxigenases de Função Mista/classificação , Oxigenases de Função Mista/genética , Oxirredução , Filogenia , Polissacarídeos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/isolamento & purificação , Especificidade por Substrato , Xilanos/química
13.
Aquat Toxicol ; 226: 105561, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32688145

RESUMO

In the aquatic environment, metals are present as mixtures, therefore studies on mixture toxicity are crucial to thoroughly understand their toxic effects on aquatic organisms. Common carp (Cyprinus carpio) were used to assess the effects of short-term Cu(II) and Cd(II) mixtures, using a fixed concentration of one of the metals, representing 25 % of its individual 96h-LC50 (concentration lethal for 50 % of the population) combined with a variable concentration of the other metal corresponding to 10, 25 or 50 % of its 96h-LC50, and vice versa. Our results showed a fast Cu and Cd bioaccumulation, with the percentage of increase in the order gill > liver > carcass. An inhibitory effect of Cu on Cd uptake was observed; higher Cu concentrations at fixed Cd levels resulted in a decreased accumulation of Cd. The presence of the two metal ions resulted in losses of total Na, K and Ca. Fish tried to compensate for the Na loss through the induction of the genes coding for Na+/K+-ATPase and H+-ATPase. Additionally, a counterintuitive induction of the gene encoding the high affinity copper transporter (CTR1) occurred, while a downregulation was expected to prevent further metal ion uptake. An induction of defensive mechanisms, both metal ion binding protein and anti-oxidant defences, was observed. Despite the metal accumulation and electrolyte loss, the low mortality suggest that common carp is able to cope with these metal levels, at least during a one-week exposure.


Assuntos
Bioacumulação/efeitos dos fármacos , Cádmio/toxicidade , Carpas/metabolismo , Cobre/toxicidade , Homeostase/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Cádmio/metabolismo , Carpas/genética , Cobre/metabolismo , Transportador de Cobre 1/genética , Transportador de Cobre 1/metabolismo , Eletrólitos/metabolismo , Brânquias/efeitos dos fármacos , Brânquias/metabolismo , Transporte de Íons , Dose Letal Mediana , Potássio/metabolismo , Sódio/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Poluentes Químicos da Água/metabolismo
14.
Nat Commun ; 11(1): 3701, 2020 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-32709883

RESUMO

Despite its importance in human cancers, including colorectal cancers (CRC), oncogenic KRAS has been extremely challenging to target therapeutically. To identify potential vulnerabilities in KRAS-mutated CRC, we characterize the impact of oncogenic KRAS on the cell surface of intestinal epithelial cells. Here we show that oncogenic KRAS alters the expression of a myriad of cell-surface proteins implicated in diverse biological functions, and identify many potential surface-accessible therapeutic targets. Cell surface-based loss-of-function screens reveal that ATP7A, a copper-exporter upregulated by mutant KRAS, is essential for neoplastic growth. ATP7A is upregulated at the surface of KRAS-mutated CRC, and protects cells from excess copper-ion toxicity. We find that KRAS-mutated cells acquire copper via a non-canonical mechanism involving macropinocytosis, which appears to be required to support their growth. Together, these results indicate that copper bioavailability is a KRAS-selective vulnerability that could be exploited for the treatment of KRAS-mutated neoplasms.


Assuntos
Neoplasias Colorretais/metabolismo , Cobre/metabolismo , Proteínas Proto-Oncogênicas p21(ras)/genética , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Animais , Disponibilidade Biológica , Sistemas CRISPR-Cas , Linhagem Celular Tumoral , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , ATPases Transportadoras de Cobre/metabolismo , Feminino , Humanos , Mucosa Intestinal/patologia , Camundongos , Camundongos Knockout , Camundongos Nus , Camundongos SCID , Mutação
15.
Proc Natl Acad Sci U S A ; 117(32): 19178-19189, 2020 08 11.
Artigo em Inglês | MEDLINE | ID: mdl-32723819

RESUMO

Lytic polysaccharide monooxygenases (LPMOs) have a unique ability to activate molecular oxygen for subsequent oxidative cleavage of glycosidic bonds. To provide insight into the mode of action of these industrially important enzymes, we have performed an integrated NMR/electron paramagnetic resonance (EPR) study into the detailed aspects of an AA10 LPMO-substrate interaction. Using NMR spectroscopy, we have elucidated the solution-phase structure of apo-BlLPMO10A from Bacillus licheniformis, along with solution-phase structural characterization of the Cu(I)-LPMO, showing that the presence of the metal has minimal effects on the overall protein structure. We have, moreover, used paramagnetic relaxation enhancement (PRE) to characterize Cu(II)-LPMO by NMR spectroscopy. In addition, a multifrequency continuous-wave (CW)-EPR and 15N-HYSCORE spectroscopy study on the uniformly isotope-labeled 63Cu(II)-bound 15N-BlLPMO10A along with its natural abundance isotopologue determined copper spin-Hamiltonian parameters for LPMOs to markedly improved accuracy. The data demonstrate that large changes in the Cu(II) spin-Hamiltonian parameters are induced upon binding of the substrate. These changes arise from a rearrangement of the copper coordination sphere from a five-coordinate distorted square pyramid to one which is four-coordinate near-square planar. There is also a small reduction in metal-ligand covalency and an attendant increase in the d(x2-y2) character/energy of the singly occupied molecular orbital (SOMO), which we propose from density functional theory (DFT) calculations predisposes the copper active site for the formation of a stable Cu-O2 intermediate. This switch in orbital character upon addition of chitin provides a basis for understanding the coupling of substrate binding with O2 activation in chitin-active AA10 LPMOs.


Assuntos
Bacillus licheniformis/enzimologia , Proteínas de Bactérias/química , Quitina/metabolismo , Oxigenases de Função Mista/química , Oxigênio/metabolismo , Bacillus licheniformis/química , Proteínas de Bactérias/metabolismo , Domínio Catalítico , Quitina/química , Cobre/química , Cobre/metabolismo , Espectroscopia de Ressonância de Spin Eletrônica , Imagem por Ressonância Magnética , Oxigenases de Função Mista/metabolismo , Oxigênio/química , Especificidade por Substrato
16.
Chemosphere ; 260: 127541, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32688311

RESUMO

The present study explores the effect of ethylene diamine disuccinic acid (EDDS) and gibberellic acid (GA) application on the phytoextraction of copper and zinc ions by Lolium perenne. When Cu was individually applied, accumulation diminished over time with little translocation from roots to shoots. In contrast, Zn accumulation and damage to roots rapidly increased over 3 days with increase in Zn translocation to shoots. Co-application of Zn to Cu amended treatments enhanced Cu concentration in shoots. For the CuEDDS application, EDDS significantly increased Cu accumulation and the damage to root increased over time, while gibberellic acid applied with Cu and Zn generally lowered metal uptake and decreased cell membrane damage. The application of EDDS and GA-EDDS, by themselves or with Cu and Zn, lowered transpiration and increased translocation, while GA increased transpiration but decreased translocation. EDDS application typically increased metal ion uptake by causing more cell damage, while GA typically lowered the damage and decreased metal uptake even though the transpiration increased over time and plant growth occurred. Furthermore, the behaviour of metal uptake changed over time and, for some treatments, the short-term and long-term response differed greatly. These results show that EDDS can be successfully used in phytoextraction of both Cu and Zn ions by Lolium perenne while GA can resist damage and protect against plant stress.


Assuntos
Biodegradação Ambiental , Etilenodiaminas/química , Giberelinas/química , Lolium/metabolismo , Poluentes do Solo/metabolismo , Transporte Biológico , Cobre/metabolismo , Etilenos , Íons/metabolismo , Raízes de Plantas/metabolismo , Succinatos/metabolismo , Zinco/metabolismo
17.
Ecotoxicol Environ Saf ; 201: 110830, 2020 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-32559689

RESUMO

Iron (Fe) oxides are intimately coupled with phosphorus and closely associated with the bioavailability of potential toxic elements (PTEs) in soil. Thus, Fe oxides may influence the stabilization of PTEs in contaminated soils amended by phosphorus. To evaluate the effects of hematite (HMT) on the stabilization of PTEs, 1-5% (by weight) of HMT was added into a contaminated red soil amended with hydroxyapatite (HAP) to simulate naturally occurring Fe oxides. The stabilization efficiencies of soil copper (Cu) and cadmium (Cd) amended with HAP in soils with low, moderate, and high content of HMT were assessed after a 60-day incubation. HAP treated the soil with high rate HMT decreased the CaCl2-extractable and acid-soluble fractions of Cu and Cd than that of HAP alone. In particular, CaCl2-extactable Cu and Cd in the soil with 5% HMT amended by HAP were 91-95% and 41-68% lower than those amended with only HAP. High content of HMT in soil could decrease the concentration of labile phosphorus in the presence of HAP, but it did not increase the concentration of NaOH-extractable inorganic phosphorus (the fraction bound to Fe oxides). The concentrations of free and crystalline Fe oxides were significantly increased by adding high dosages of HMT with or without HAP. High content of HMT in soil amended by HAP reduced metal phytotoxicity and uptake by wheat shoots than the soil containing HAP without HMT. The results indicate that HMT can promote Cu and Cd stabilization while decrease labile phosphorus in red soil amended with HAP, suggesting that phosphorus-based amendments combined with Fe oxides can be used to stabilize PTEs in contaminated red soils.


Assuntos
Cádmio/análise , Cobre/análise , Durapatita/química , Compostos Férricos/química , Fósforo/análise , Poluentes do Solo/análise , Disponibilidade Biológica , Cádmio/metabolismo , China , Cobre/metabolismo , Fósforo/metabolismo , Solo/química , Poluentes do Solo/metabolismo , Triticum/efeitos dos fármacos , Triticum/metabolismo
18.
Chemosphere ; 259: 127436, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32599387

RESUMO

Copper (Cu) is an essential metal for human, animals and plants, although it is also potentially toxic above supra-optimal levels. In plants, Cu is an essential cofactor of numerous metalloproteins and is involved in several biochemical and physiological processes. However, excess of Cu induces oxidative stress inside plants via enhanced production of reactive oxygen species (ROS). Owing to its dual nature (essential and a potential toxicity), this metal involves a complex network of uptake, sequestration and transport, essentiality, toxicity and detoxification inside the plants. Therefore, it is vital to monitor the biogeo-physiochemical behavior of Cu in soil-plant-human systems keeping in view its possible essential and toxic roles. This review critically highlights the latest understanding of (i) Cu adsorption/desorption in soil (ii) accumulation in plants, (iii) phytotoxicity, (iv) tolerance mechanisms inside plants and (v) health risk assessment. The Cu-mediated oxidative stress and resulting up-regulation of several enzymatic and non-enzymatic antioxidants have been deliberated at molecular and cellular levels. Moreover, the role of various transporter proteins in Cu uptake and its proper transportation to target metalloproteins is critically discussed. The review also delineates Cu build-up in plant food and accompanying health disorders. Finally, this review proposes some future perspectives regarding Cu biochemistry inside plants. The review, to a large extent, presents a complete picture of the biogeo-physiochemical behavior of Cu in soil-plant-human systems supported with up-to-date 10 tables and 5 figures. It can be of great interest for post-graduate level students, scientists, industrialists, policymakers and regulatory authorities.


Assuntos
Cobre/metabolismo , Poluentes do Solo/metabolismo , Antioxidantes/metabolismo , Cobre/toxicidade , Humanos , Estresse Oxidativo/fisiologia , Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Medição de Risco , Solo/química , Poluentes do Solo/química , Poluentes do Solo/toxicidade
19.
Nucleic Acids Res ; 48(13): 7356-7370, 2020 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-32520335

RESUMO

To enable the optimal, biocompatible and non-destructive application of the highly useful copper (Cu+)-mediated alkyne-azide 'click' cycloaddition in water, we have isolated and characterized a 79-nucleotide DNA enzyme or DNAzyme, 'CLICK-17', that harnesses as low as sub-micromolar Cu+; or, surprisingly, Cu2+ (without added reductants such as ascorbate) to catalyze conjugation between a variety of alkyne and azide substrates, including small molecules, proteins and nucleic acids. CLICK-17's Cu+ catalysis is orders of magnitude faster than that of either Cu+ alone or of Cu+ complexed to PERMUT-17, a sequence-permuted DNA isomer of CLICK-17. With the less toxic Cu2+, CLICK-17 attains rates comparable to Cu+, under conditions where both Cu2+ alone and Cu2+ complexed with a classic accelerating ligand, THPTA, are wholly inactive. Cyclic voltammetry shows that CLICK-17, unlike PERMUT-17, powerfully perturbs the Cu(II)/Cu(I) redox potential. CLICK-17 thus provides a unique, DNA-derived ligand environment for catalytic copper within its active site. As a bona fide Cu2+-driven enzyme, with potential for being evolved to accept only designated substrates, CLICK-17 and future variants promise the fast, safe, and substrate-specific catalysis of 'click' bioconjugations, potentially on the surfaces of living cells.


Assuntos
Cobre/metabolismo , Reação de Cicloadição/métodos , DNA Catalítico/química , Alquinos/química , Azidas/química , Química Click/métodos , Oxirredução , Água/química
20.
Bull Environ Contam Toxicol ; 104(6): 755-762, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32394051

RESUMO

We examined copper accumulation in the hemolymph, gills and hepatopancreas, and hemolymph osmolality, Na+ and Cl- concentrations, together with gill Na+/K+-ATPase and carbonic anhydrase activities, after dietary copper delivery (0, 100 or 500 Cu µg g-1) for 12 days in a fiddler crab, Minuca rapax. In contaminated crabs, copper concentration decreased in the hemolymph and hepatopancreas, but increased in the gills. Hemolymph osmolality and gill Na+/K+-ATPase activity increased while hemolymph [Na+] and [Cl-] and gill carbonic anhydrase activity decreased. Excretion likely accounts for the decreased hemolymph and hepatopancreas copper titers. Dietary copper clearly affected osmoregulatory ability and hemolymph Na+ and Cl- regulation in M. rapax. Gill copper accumulation decreased carbonic anhydrase activity, suggesting that dietary copper affects acid-base balance. Elevated gill Na+/K+-ATPase activity appears to compensate for the ion-regulatory disturbance. These effects of dietary copper illustrate likely impacts on semi-terrestrial species that feed on metal-contaminated sediments.


Assuntos
Bioacumulação , Braquiúros/efeitos dos fármacos , Cobre/metabolismo , Osmorregulação/efeitos dos fármacos , Poluentes Químicos da Água/metabolismo , Animais , Braquiúros/metabolismo , Cobre/análise , Exposição Dietética , Brânquias/efeitos dos fármacos , Brânquias/metabolismo , Hemolinfa/metabolismo , Hepatopâncreas/metabolismo , Transporte de Íons , Concentração Osmolar , Alimentos Marinhos , ATPase Trocadora de Sódio-Potássio/metabolismo , Poluentes Químicos da Água/análise
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