Trends in metal-binding and metalloprotein analysis

Garcia, Jerusa Simone; Magalhães, Cristiana Schmidt de; Arruda, Marco Aurélio Zezzi

doi:10.1016/j.talanta.2005.08.041

Cited by 149 publications

(125 citation statements)

References 61 publications

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“…Overall, the above-mentioned papers document the feasibility and the effectiveness of a classic proteomic approach. Moreover, it is worth noting here that in the case of metallodrugs additional proteomic experiments may be devised that are centered on the metal and are principally aimed at investigating metal distribution patterns within the proteome (metallomics) [22][23][24]. These investigative approaches, relying on the selective tagging of the metallodrug or of the metal itself within 1D or 2D gels through highly sensitive methods, allow in principle the identification of all those proteins capable of binding the metallodrug of interest.…”

Section: Introductionmentioning

confidence: 99%

Exploring the biochemical mechanisms of cytotoxic gold compounds: a proteomic study

et al. 2010

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We have recently shown that a group of structurally diverse gold compounds are highly cytotoxic toward a panel of 36 human tumor cell lines through a variety of biochemical mechanisms. A classic proteomic approach is exploited here to gain deeper insight into those mechanisms. This investigation is focused on Auoxo6, a novel binuclear gold(III) complex, and auranofin, a clinically established gold(I) antiarthritic drug. First, the 72-h cytotoxicity profiles of Auoxo6 and auranofin were determined against A2780 human ovarian carcinoma cells. Subsequently, protein extraction from gold-treated A2780 cells sensitive to cisplatin and 2D gel electrophoresis separation were carried out according to established procedures. Notably, both metallodrugs caused relatively modest changes in protein expression in comparison with controls as only 11 out of approximately 1,300 monitored spots showed appreciable quantitative changes. Very remarkably, six altered proteins were in common between the two treatments. Eight altered proteins were identified by mass spectrometry; among them was ezrin, a protein associated with the cytoskeleton and involved in apoptosis. Interestingly, two altered proteins, i.e., peroxiredoxins 1 and 6, are known to play crucial roles in the cell redox metabolism. Increased cleavage of heterogeneous ribonucleoprotein H was also evidenced, consistent with caspase 3 activation. Overall, the results of the present proteomic study point out that the mode of action of Auoxo6 is strictly related to that of auranofin, that the induced changes in protein expression are limited and selective, that both gold compounds trigger caspase 3 activation and apoptosis, and that a few affected proteins are primarily involved in cell redox homeostasis.

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Section: Introductionmentioning

confidence: 99%

Exploring the biochemical mechanisms of cytotoxic gold compounds: a proteomic study

et al. 2010

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“…A recent review describes the state-of-art, tendencies and analysis related to the metal-binding proteins and metalloproteins. 5 Metal-binding proteins and metalloproteins represent a large portion of the number of proteins. It has been estimated that around 40% of all proteins and enzymes contain metal ions in their structures.…”

Section: Introductionmentioning

confidence: 99%

“…As a consequence, they are found in the majority of metalloproteins. 5 Fruits are one source of metalloproteins, where the elements are mainly associated to albumins and globulins. 3 The most common means to accomplish element speciation consist of combined and/or coupled systems, in which species are separated before the selective detection of the elements.…”

Section: Introductionmentioning

confidence: 99%

“…6 The coupling of chromatography by using different separation mechanisms with inductively coupled plasma mass spectrometry (ICP-MS) detection has been the most widely used analytical tool for element speciation. [5][6][7][8] In the latest years, fruits and nuts from Amazon region have arisen interest among the scientific community due to the high concentrations of some essential elements in their compositions. [9][10][11][12][13][14][15][16][17] Special attention are devoted to Brazil-nut (Bertholletia excelsa), known as "castanha-doPará".…”

Section: Introductionmentioning

confidence: 99%

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Cu, Fe, Mn and Zn distribution in protein fractions of Brazil-nut, cupuassu seed and coconut pulp by solid-liquid extraction and electrothermal atomic absorption spectrometry

Naozuka¹,

Oliveira²

2007

J. Braz. Chem. Soc.

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Neste trabalho foram determinados Cu, Fe, Mn and Zn em extratos de castanha-do-Pará (Bertholletia excelsa), semente de cupuaçu (Theobroma grandiflorum) e polpa de coco (Cocos nucifera) usando extração sólido-líquido e espectrometria de absorção atômica com atomização eletrotérmica. Foram empregadas diferentes soluções extratoras (mistura de metanol e clorofórmio 1:2 v/v, água, 0,5 mol L -1 NaCl, 70% v/v etanol e 0,5 mol L -1 NaOH) para extração de lipídios, albuminas, globulinas, prolaminas e glutelinas, respectivamente. As concentrações de proteínas encontradas na castanha-do-pará foram maiores do que aquelas encontradas na semente de cupuaçu e polpa de coco. Nas frações lipídicas foram encontradas maiores concentrações de Fe em castanhado-pará e semente de cupuaçu e Zn em polpa de coco. As determinações dos elementos nos extratos proteicos de castanha-do-pará indicaram que eles estão ligados, preferencialmente, às albuminas, globulinas e glutelinas. O mesmo comportamento não foi observado para semente de cupuaçu e polpa de coco. As concetrações dos elementos variaram para os diferentes extratos proteícos. Em geral, as maiores concentrações de proteínas foram determinadas nos extratos em NaOH. Por outro lado, as menores concentrações de Cu, Fe, Mn e Zn foram encontradas nessa fração.In this work Cu, Fe, Mn and Zn were determined in extracts of Brazil-nut (Bertholletia excelsa), cupuassu (Theobroma grandiflorum) seeds and coconut (Cocos nucifera) pulp, using solid-liquid extraction and electrothermal atomic absorption spectrometry. Different extractant solutions (mixture of methanol and chloroform 1:2 v/v, water, 0.5 mol L -1 NaCl, 70% v/v ethanol, and 0.5 mol L -1 NaOH) were used for extracting lipids, albumins, globulins, prolamins and glutelins, respectively. The protein concentrations determined in Brazil-nut are higher than those observed for cupuassu and coconut pulp. The analysis of lipid fractions reveals high concentations of Fe in Brazil-nut and cupuassu seed and Zn in coconut pulp. For Brazil-nut, these elements are mainly associated to albumin, globulin and glutelin fractions. The same behaviour was not observed for cupuassu seed and coconut pulp. The element concentrations changed in each protein fraction. In general, high concentrations of proteins were determined in NaOH extractants. On the other hand, low concentration of Cu, Fe, Mn and Zn were found in this fraction.Keywords: solid-liquid extraction, electrothermal atomic absorption spectrometry, Brazilnut, cupuassu, coconut IntroductionFoods are one of the main sources of metals for human body. Most of the metal ions present in foods are bound to specific proteins or enzymes and exert their effects as active or structural centers, command process such as catalysts, substrate binding and activation, transportation and storage. 1,2 In this way, the knowledge of the foods compositional, nutritional and functional properties is fundamental for defining their quality. In general, nutritional properties are characterized by both the abundance and bio...

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“…O estudo da metalômica das diferentes espécies de peixes poderá fornecer informações valiosas sobre como um íon metálico está distribuído e coordenado às proteínas, como também da concentração individual da espécie metálica, contribuindo assim na elucidação dos aspectos fisiológicos e funcionais dessas biomoléculas. [10][11][12][13] Recentemente foram publicados por pesquisadores brasileiros os primeiros resultados obtidos no estudo metaloproteômico da espécie de peixe tilápia do Nilo. 14,15 Nessas publicações, os autores, utilizando 2D-PAGE no fracionamento das proteínas de plasma e tecido hepático, identificaram, por SR XRF, manganês e zinco em 10 proteínas do plasma e cálcio, ferro e zinco em 26 proteínas do fígado.…”

Section: Introductionunclassified

Fracionamento de cobre em proteínas do plasma, músculo e fígado de tilápia do Nilo

Neves¹,

Lima²,

Baldassini³

et al. 2012

Quím. Nova

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Recebido em 18/4/11; aceito em 7/9/11; publicado na web em 3/11/11 COPPER FRACTIONATION IN PROTEINS FROM PLASMA, MUSCLE AND LIVER OF NILE TILAPIA. Copper fractionation in plasma, muscle and liver of Nile tilapia was performed after protein separation by 2D-PAGE. SR XRF analysis indicated the presence of copper in three protein spots of plasma, and in two protein spots of muscle and liver, respectively. Copper ions were found to be distributed mostly in proteins that had a molar mass of less than 54 kDa and greater than 13 kDa and a pI in the 5.3-9.3 range. The copper concentration bound to these proteins was determined by GFAAS which showed concentrations in the 1.20-4.82 mg g -1 range.Keywords: Nile tilapia; metalloproteins; metallomics. INTRODUÇÃONa nutrição de peixes, como de outros animais, o cobre é elemento dietético essencial. Tanto a deficiência quanto o excesso desse mineral podem causar alterações nas funções fisiológicas e na resposta imunológica, aumentando a susceptibilidade do animal a infecções.1,2 Esse mineral possui importantes funções no organismo dos peixes, sendo necessário no metabolismo de vários nutrientes e como constituinte de diversas enzimas. Destaca-se como referência de sua importância a participação como cofator, dentre outras, da enzima ALA-desidrogenase, a qual participa na síntese do heme em sua incorporação no pigmento hemoglobina. Além disso, é indiretamente responsável também pela formação do colágeno e da elastina, e pela conversão da tirosina em melanina em destruição de ânions superóxido, que são espécies reativas do oxigênio, potencialmente destrutivos para as células. [1][2][3][4] A importância dos minerais como componentes estruturais e funcionais dos organismos vivos e o fato de grande parte das proteínas e enzimas requererem a presença de um íon metálico para realizar alguma atividade biológica levaram os pesquisadores que trabalham com aquicultura a desenvolverem trabalhos relacionados à influência das metaloproteínas na fisiologia, genética e nutrição de peixes. [4][5][6][7][8][9] Nesse contexto, atualmente, o desenvolvimento de metodologias analíticas que permitam uma avaliação segura das metaloproteínas e/ou proteínas ligadas a metais presentes no sangue e em tecidos de animais se tornou fundamental. Principalmente, em relação aos chamados microminerais, como o cobre, os quais apresentam faixas de essencialidade e toxicidade muito próximas e que são transportados em nível celular por diferentes proteínas. Considerando a importância que essas biomoléculas exercem sobre as atividades biológicas dos seres vivos, uma nova área científica denominada metalômica foi proposta recentemente e permitiu a integração de estudos tradicionalmente analíticos com estudos inorgânicos e bioquímicos. O estudo da metalômica das diferentes espécies de peixes poderá fornecer informações valiosas sobre como um íon metálico está distribuído e coordenado às proteínas, como também da concentração individual da espécie metálica, contribuindo assim na elucidação dos aspectos fisiológicos e fun...

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Trends in metal-binding and metalloprotein analysis

Cited by 149 publications

References 61 publications

Exploring the biochemical mechanisms of cytotoxic gold compounds: a proteomic study

Exploring the biochemical mechanisms of cytotoxic gold compounds: a proteomic study

Cu, Fe, Mn and Zn distribution in protein fractions of Brazil-nut, cupuassu seed and coconut pulp by solid-liquid extraction and electrothermal atomic absorption spectrometry

Fracionamento de cobre em proteínas do plasma, músculo e fígado de tilápia do Nilo

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