In recent decades, the scientific community has widely debated the contamination of fish in the Amazon region by mercury species. As the diet of riverside populations in the Amazon region is based mainly on fish, these populations are exposed to mercurial species that can cause serious and irreversible damage to their health. The risks of consuming fish exposed to mercurial species in the Amazon region have motivated toxicological investigations. However, the effect of mercurial species on protein and enzyme levels is still controversial. In this work, analytical and bioanalytical techniques Two-dimensional polyacrylamide gel electrophoresis [2D-PAGE] Graphite Furnace Atomic Absorption Spectrometry [GFAAS], and Mass Spectrometry in Sequence with Electrospray Ionization [ESI–MS/MS] were used to identify proteins associated with mercury (metal-binding protein) in muscle and liver tissues of the fish species Pinirampus pirinampu from the Madeira River, in the Brazilian Amazon. Enzymatic and lipid peroxidation analyses were also used to assess changes related to oxidative stress. Determinations of total mercury by GFAAS indicated higher concentrations in liver tissue (555 ± 19.0 µg kg⁻¹) when compared to muscle tissue (60 ± 2.0 µg kg⁻¹). The fractionation process of tissue proteomes by 2D-PAGE and subsequent mapping of mercury by GFAAS in the protein spots of the gels identified the presence of mercury in three spots of the liver tissue (concentrations in the range of 0.800 to 1.90 mg kg⁻¹). The characterization of protein spots associated with mercury by ESI–MS/MS identified the enzymes triosephosphate isomerase A, adenylate kinase 2 mitochondrial, and glyceraldehyde-3-phosphate dehydrogenase as possible candidates for mercury exposure biomarkers. The muscle tissue did not show protein spots associated with mercury. Enzymatic activity decreased proportionally to the increase in mercury concentrations in the tissues.

Graphical abstract

中文翻译：

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巴西亚马孙河马德拉河中 Pinirampus pirinampu 暴露于汞物种中蛋白质生物标志物和氧化应激的研究

近几十年来，科学界广泛讨论了亚马逊地区鱼类被汞污染的问题。由于亚马逊地区沿河人口的饮食主要以鱼类为食，因此这些人口接触到汞的物种，可能对其健康造成严重且不可逆转的损害。在亚马逊地区食用暴露于汞物种的鱼类的风险激发了毒理学研究。然而，汞对蛋白质和酶水平的影响仍然存在争议。在这项工作中，分析和生物分析技术 二维聚丙烯酰胺凝胶电泳 [2D-PAGE] 石墨炉原子吸收光谱 [GFAAS]，来自巴西亚马逊马德拉河的Pinirampus pirinampu 。酶促和脂质过氧化分析也用于评估与氧化应激相关的变化。^{与肌肉组织 (60 ± 2.0 µg kg -1} )相比，通过 GFAAS 对总汞的测定表明肝组织中的汞浓度 (555 ± 19.0 µg kg ^-1 ) 更高。通过 2D-PAGE 对组织蛋白质组的分离过程以及随后通过 GFAAS 对凝胶蛋白质点中的汞作图确定了在肝脏组织的三个点中存在汞（浓度范围为 0.800 至 1.90 mg kg ^-1） . 蛋白质斑点的表征通过 ESI-MS/MS 与汞相关的酶确定了磷酸丙糖异构酶 A、腺苷酸激酶 2 线粒体和 3-磷酸甘油醛脱氢酶作为汞暴露生物标志物的可能候选者。肌肉组织没有显示出与汞相关的蛋白质斑点。酶活性与组织中汞浓度的增加成比例下降。

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