Metallomic study on plasma samples from Nile tilapia using SR-XRF and GFAAS after separation by 2D PAGE: initial results

Santos, Felipe A.; Lima, Paula Monteiro de; Neves, R. F.; Moraes, Paula Martin de; Pérez, Carlos A.; Silva, Marcelo O. A.; Arruda, Marco Aurélio Zezzi; Castro, Gustavo Rocha de; Padilha, Pedro M.

doi:10.1007/s00604-010-0522-y

Cited by 26 publications

(26 citation statements)

References 20 publications

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“…For 2D-PAGE separation, only the pellets from the second precipitation (Mw \ 90 kDa) were used. The two electrophoretic steps were performed as described previously (Lima et al 2010;Santos et al 2011;Neves et al 2012;Silva et al 2013).…”

Section: Electrophoretic Runs (2d-page)mentioning

confidence: 99%

“…This binding process can inactivate enzymes and alter cell activity resulting in the loss of skin sensitivity and muscle coordination, deafness, and death (Bakir et al 1973;Clarkson 1990;Pollard and Hultman 1997;Jesus and Carvalho 2008). The aim of our research group was to perform a metalloproteomic study (Santos et al 2011;Moraes et al 2012;Braga et al 2015) for separate twodimensional polyacrylamide gel electrophoresis (2D-PAGE) determinations to determine the quantity of Hg in the protein spots (using graphite furnace atomic absorption spectrometry [GFAAS]) to characterize proteins bound to Hg (using electrospray ionization tandem mass spectrometry [ESI-MS/MS]) and to identify potential biomarkers of metalloproteins in muscle and liver tissue samples from Tucunaré (Cichla spp.) collected in the Amazon region (from hydroelectric dams).…”

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confidence: 99%

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Determination of the Mercury Fraction Linked to Protein of Muscle and Liver Tissue of Tucunaré (Cichla spp.) from the Amazon Region of Brazil

Vieira

Cavecci

Queiroz

et al. 2015

Arch Environ Contam Toxicol

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This study used metalloproteomic techniques to characterize mercury (Hg)-bound proteins in the muscle and liver tissue of Tucunaré (Cichla spp.) collected at the Jirau Hydroelectric Power Plant in Madeira River Basin, Brazil. The proteome of the muscle and liver tissue was obtained after two steps of fractional precipitation and separating the proteins by 2-D polyacrylamide gel electrophoresis. Hg was identified and quantified in the protein spots by graphite furnace atomic absorption spectrometry after acid mineralization in an ultrasound bath. Hg with a molecular weight <20 kDa and a concentration between 13.30 and 33.40 mg g(-1) was found in the protein spots. These protein spots were characterized by electrospray ionization tandem mass spectrometry after trypsin digestion. From a total of 12 analyzed spots, seven proteins showing Hg biomarker characteristics were identified: parvalbumin and its isoforms, ubiquitin-40S ribosomal protein S27a, zinc (Zn) finger and BTB domain-containing protein 24, and dual-specificity protein phosphatase 22-B.

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Section: Electrophoretic Runs (2d-page)mentioning

confidence: 99%

mentioning

confidence: 99%

Determination of the Mercury Fraction Linked to Protein of Muscle and Liver Tissue of Tucunaré (Cichla spp.) from the Amazon Region of Brazil

Vieira

Cavecci

Queiroz

et al. 2015

Arch Environ Contam Toxicol

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“…The correlation analysis of the gels demonstrated that 68% and 70% of the protein spots were present in the four gels of the dourada and pacu muscle tissue samples, respectively. In addition, the average number of spots found in the gels -168±15 in dourada and 98±8 in pacu -presented a relative standard deviation of less than 10%, which is considered very good for protein fractionation by 2D-PAGE (Lima et al, 2010;Santos et al, 2011).…”

Section: Resultsmentioning

confidence: 99%

Metallomic study of mercury in fish from Amazon region - Brazil

Moraes

Santos

Padilha

et al. 2013

E3S Web of Conferences

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Abstract. This paper presents preliminary findings for a metallomics study of mercury in the muscle of the fish from Amazonas region -Brazil, after protein separation by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and subsequent evaluation of mercury by Thermal Decomposition and Amalgamation Coupled with Atomic Absorption (TTA-CAAS). It was found that mercury is present in 18 protein spots of "dourada" muscle (Brachyplatystoma rousseauxii) and 9 protein spots of "pacu" muscle (Myleus sp.). The protein spots in which they were determined the presence of mercury present molecular weight of 13.5 and 21.4 kDa and isoelectric point of 3.8 and 9.2, respectively.

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“…O clima na bacia do rio Solimões, é bem semelhante ao da bacia amazônica, sendo quente e úmido, com precipitação pluvial anual igual ou superior a 2.000 mm e com médias anuais de temperatura e umidade relativa de 27 °C e 84%, respectivamente (GUIMARÃES et al, 2013). Essas fitas foram colocadas na caixa de hidratação IPGBOX, cobertas com óleo mineral e mantidas à temperatura ambiente por aproximadamente 12 h. Após esse período, as fitas foram transferidas para o isoeletrofocalizador EttanTM IPGphorTM 3 para realização da primeira dimensão do processo eletroforético (LIMA et al, 2010;NEVES et al, 2012;SANTOS et al, 2011;SILVA et al, 2013).…”

Section: Rio Apuaúunclassified

“…O corante ficou em contato com o gel durante 72 h em mesa agitadora a 40 rpm e depois foi removido através de sucessivas lavagens com água ultrapura até que as proteínas (spots) ficassem visíveis (LIMA et al, 2010;NEVES et al, 2012;SANTOS et al, 2011;SILVA et al, 2013 Já a estatística descritiva foi utilizada para apresentar características gerais (comprimento, peso e concentração de mercúrio total) dos tucunarés por local de amostragem.…”

Section: Rio Purusunclassified

Estratégias metaloproteômicas na investigação de biomarcadores de mercúrio em amostras de tecido muscular de tucunarés

Figueiredo¹

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Formação acadêmica -2012Graduação em Gestão Ambiental. Universidade de Brasília, UnB, Brasil total no tecido muscular de tucunarés capturados em rios e reservatórios hidrelétricos.As concentrações apresentaram pouca variação independente do local de amostragem e algumas estavam superiores aos limites máximos definidos na legislação brasileira. As concentrações aumentaram na sequência: reservatório de Cana Brava < reservatório de Balbina < rio Madeira < rio Purus < reservatório de Tucuruí < rio Apuaú < rio Solimões < rio Negro. As amostras do rio Negro foram selecionadas para definição de um biomarcador proteico através da eletroforese bidimensional (2D-PAGE). O mercúrio foi determinado em 15 possíveis proteínas com massa molar abaixo de 20,1 kDa, sendo 2 mais promissoras à biomarcadoras. Além disso, foi extraído da musculatura dos tucunarés apenas 12,4% do mercúrio, sendo que 89% desse mercúrio foi aplicado na técnica 2D-PAGE, resultando em 49,74% de mercúrio nas proteínas. Essa porcentagem de mercúrio nos spots indica que os procedimentos otimizados de preparação da amostra foram eficientes para ser aplicados na 2D-PAGE. Os resultados metaloproteômicos desse estudo são importantes no subsídio de estudos futuros de identificação e caracterização de biomarcadores de mercúrio em peixes.Descritores: biomarcadores; metaloproteínas; contaminantes; peixes. Thus, this study sought to identify possible mercury exposure biomarkers in tucunares the Negro river through metalproteomics methods. Initially, were determined the total mercury concentrations in muscle tissue of tucunares caught in rivers and hydroelectric reservoirs. They were in a little independent variation of the sampling site and some were above the maximum limits set by Brazilian law. The concentrations increased following: Cana Brava reservoir < Balbina reservoir < Madeira river < Purus river < Tucurui reservoir < Apuau river < Solimões river < Negro river. Samples of the Negro river were selected for definition of a protein biomarker by two-dimensional electrophoresis (2D-PAGE). Mercury was determined in 15 possible protein with molecular weight of 20.1 kDa below, 2 the most promising biomarkers. Moreover, it was extracted from the muscle of tucunarés only 12.4% of mercury, with 89% of mercury was applied to the 2D-PAGE technique, and 49.74% of mercury in proteins.This percentage of mercury in spots indicates that the sample preparation procedures were optimized for efficient be applied to 2D-PAGE. The metalproteomics results of this study are important in the allowance for futures studies of identification and characterization of mercury biomarkers in fish.

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Metallomic study on plasma samples from Nile tilapia using SR-XRF and GFAAS after separation by 2D PAGE: initial results

Cited by 26 publications

References 20 publications

Determination of the Mercury Fraction Linked to Protein of Muscle and Liver Tissue of Tucunaré (Cichla spp.) from the Amazon Region of Brazil

Determination of the Mercury Fraction Linked to Protein of Muscle and Liver Tissue of Tucunaré (Cichla spp.) from the Amazon Region of Brazil

Metallomic study of mercury in fish from Amazon region - Brazil

Estratégias metaloproteômicas na investigação de biomarcadores de mercúrio em amostras de tecido muscular de tucunarés

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