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利用金属蛋白质组学策略鉴定亚马逊河圆口脂鲤中汞暴露的生物标志物蛋白

Metalloproteomic Strategies for Identifying Proteins as Biomarkers of Mercury Exposure in Serrasalmus rhombeus from the Amazon Region.

机构信息

School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Rua Prof R. Dr. Valter Maurício Corrêa, s/n, Botucatu, SP, 18.618-681, Brazil.

Venom and Venomous Animals Study Center (CEVAP) of São Paulo State University (UNESP), Rua José Barbosa de Barros, 1780 - Vila Ema, Botucatu, SP, 18.610-307, Brazil.

出版信息

Biol Trace Elem Res. 2021 Feb;199(2):712-720. doi: 10.1007/s12011-020-02178-9. Epub 2020 May 24.

Abstract

This manuscript describes the results of a metalloproteomic study of mercury in samples of muscle and liver tissue of the species Serrasalmus rhombeus, popularly known as black piranha and characterised as the most voracious and aggressive predator in the Brazilian Amazon. The metalloproteomic study involved using two-dimensional electrophoresis (2D PAGE) to fractionate the proteome of the muscle and liver tissue samples, along with atomic absorption spectrometry in a graphite furnace (GFAAS) to identify mercury associated with protein SPOTs and mass spectrometry with electrospray ionisation (ESI-MS/MS) to characterise the mercury-binding proteins. The protein SPOTs characterised showed concentrations in the order of 156 mg kg, which ranks as the highest concentrations of mercury determined so far in metalloproteomic studies involving fish species in the Amazon region. Based on FASTA sequences of proteins characterised by ESI-MS/MS, bioinformatics studies were performed that allowed identifying nine proteins with characteristics of biomarkers of mercury exposure. Of those proteins, glutathione peroxidase stands out as an enzyme of great importance in the antioxidant defence of organisms subjected to oxidative stress caused by xenobiotics.

摘要

这篇手稿描述了一项关于汞的金属蛋白质组学研究结果,该研究的样本来自罗贝氏锯脂鲤(俗称黑食人鱼)的肌肉和肝脏组织。罗贝氏锯脂鲤是巴西亚马逊地区最贪婪、最具攻击性的捕食者。金属蛋白质组学研究涉及使用二维电泳(2D PAGE)对肌肉和肝脏组织样本的蛋白质组进行分离,同时使用石墨炉原子吸收光谱法(GFAAS)来鉴定与蛋白质斑点结合的汞,以及使用电喷雾电离(ESI-MS/MS)的质谱法来对结合汞的蛋白质进行特征分析。所鉴定的蛋白质斑点的浓度顺序为 156 毫克/千克,这是迄今为止在涉及亚马逊地区鱼类的金属蛋白质组学研究中确定的最高汞浓度。基于 ESI-MS/MS 鉴定的蛋白质的 FASTA 序列,进行了生物信息学研究,从而确定了 9 种具有汞暴露生物标志物特征的蛋白质。在这些蛋白质中,谷胱甘肽过氧化物酶作为一种酶,在生物体的抗氧化防御中具有重要作用,可对抗由外源性化学物质引起的氧化应激。

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