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利用金属蛋白质组学策略鉴定巴西马德拉河圆口铜鱼汞污染的生物标志物。

Identification of Biomarkers of Mercury Contamination in Brachyplatystoma filamentosum of the Madeira River, Brazil, Using Metalloproteomic Strategies.

机构信息

School of Veterinary Medicine and Animal Science, São Paulo State University (UNESP), Botucatu, Brazil.

Institute of Biosciences, São Paulo State University (UNESP), Botucatu, Brazil.

出版信息

Biol Trace Elem Res. 2019 Jan;187(1):291-300. doi: 10.1007/s12011-018-1363-5. Epub 2018 May 8.

DOI:10.1007/s12011-018-1363-5
PMID:29740802
Abstract

Predator fish can accumulate high levels of mercury, which qualifies them as potential indicators of this toxic metal. The predatory species Brachyplatystoma filamentosum, popularly known as filhote, is among the most consumed species in the Brazilian Amazon. Continuing the metalloproteomic studies of mercury in Amazonian fishes that have been developed in the last 5 years, the present paper provides the data of protein characterization associated with mercury in muscle and liver samples of filhote (Brachyplatystoma filamentosum) collected in the Madeira River, Brazilian Amazon. The mercury concentration in the muscle and liver samples was determined by graphite furnace atomic absorption spectrometry (GFAAS). The protein fraction was extracted in an aqueous medium, and later, a fractional precipitation procedure was performed to obtain the protein pellets. Then, the proteome of the tissue samples of this fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and a mercury mapping of the protein spots was carried out by GFAAS after acid digestion. Protein spots that had mercury were characterized by mass spectrometry with electrospray ionization in sequence (ESI-MS/MS) after tryptic digestion. It was possible to characterize 11 mercury-associated protein spots that presented biomarker characteristics and could be used to monitor mercury in fish species of the Amazon region. Thus, the metalloproteomic strategies used in the present study allowed us to characterize 11 mercury-associated protein spots. It should be noted that the protein spots identified as GFRP, TMEM186, TMEM57B, and BHMT, which have coordination sites for elements with characteristics of soft acids, such as mercury, can be used as biomarkers of mercury contamination in monitoring studies of this toxic metal in fish species from the Amazon region.

摘要

掠食性鱼类可能会积累大量的汞,这使它们成为这种有毒金属的潜在指标。在巴西亚马逊地区,最受欢迎的掠食性物种之一是 Brachyplatystoma filamentosum,俗称 filhote。在过去 5 年中,对亚马逊鱼类中汞的金属蛋白质组学研究不断进行,本研究提供了在马德拉河(巴西亚马逊地区)采集的 filhote(Brachyplatystoma filamentosum)肌肉和肝脏样本中与汞相关的蛋白质特征的数据。肌肉和肝脏样本中的汞浓度通过石墨炉原子吸收光谱法(GFAAS)测定。在水介质中提取蛋白质馏分,然后进行分级沉淀程序以获得蛋白质沉淀。然后,通过二维聚丙烯酰胺凝胶电泳(2D-PAGE)分离该鱼类组织样本的蛋白质组,并在酸消化后通过 GFAAS 对蛋白质斑点进行汞映射。通过电喷雾电离序列(ESI-MS/MS)对经胰蛋白酶消化后含有汞的蛋白质斑点进行质谱分析。共鉴定出 11 个具有生物标志物特征的与汞相关的蛋白质斑点,可用于监测亚马逊地区鱼类中的汞。因此,本研究中使用的金属蛋白质组学策略使我们能够鉴定 11 个与汞相关的蛋白质斑点。值得注意的是,鉴定出的 GFRP、TMEM186、TMEM57B 和 BHMT 等蛋白质斑点具有与软酸(如汞)特性相似的配位位点,可作为监测亚马逊地区鱼类中这种有毒金属污染的生物标志物。

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