Institute of Bioscience of Botucatu, São Paulo State University (UNESP), Rubião Júnior, Botucatu, São Paulo, 18618-970, Brazil.
Pontifical Catholic University of Goiás (PUC), Goiânia, GO, Brazil.
Biol Trace Elem Res. 2018 May;183(1):164-171. doi: 10.1007/s12011-017-1129-5. Epub 2017 Aug 21.
This study presents data on the extraction and characterization of proteins associated with mercury in the muscle and liver tissues of jaraqui (Semaprochilodus spp.) from the Madeira River in the Brazilian Amazon. Protein fractionation was carried out by two-dimensional electrophoresis (2D-PAGE). Mercury determination in tissues, pellets, and protein spots was performed by graphite furnace atomic absorption spectrometry (GFAAS). Proteins in the spots that showed mercury were characterized by electrospray ionization tandem mass spectrometry (ESI-MS/MS). The highest mercury concentrations were found in liver tissues and pellets (426 ± 6 and 277 ± 4 μg kg), followed by muscle tissues and pellets (132 ± 4 and 86 ± 1 μg kg, respectively). Mercury quantification in the protein spots allowed us to propose stoichiometric ratios in the range of 1-4 mercury atoms per molecule of protein in the protein spots. The proteins characterized in the analysis by ESI-MS/MS were keratin, type II cytoskeletal 8, parvalbumin beta, parvalbumin-2, ubiquitin-40S ribosomal S27a, 39S ribosomal protein L36 mitochondrial, hemoglobin subunit beta, and hemoglobin subunit beta-A/B. The results suggest that proteins such as ubiquitin-40S ribosomal protein S27a, which have specific domains, possibly zinc finger, can be used as biomarkers of mercury, whereas mercury and zinc present characteristics of soft acids.
本研究提供了关于从巴西亚马逊马德拉河中提取并鉴定与汞相关的肌肉和肝脏组织蛋白的数据。采用二维电泳(2D-PAGE)进行蛋白质分级分离。用石墨炉原子吸收光谱法(GFAAS)测定组织、沉淀和蛋白点中的汞含量。通过电喷雾串联质谱(ESI-MS/MS)对显示汞的蛋白点进行鉴定。肝组织和沉淀中的汞浓度最高(426±6 和 277±4μg/kg),其次是肌肉组织和沉淀(分别为 132±4 和 86±1μg/kg)。对蛋白点中汞的定量分析使我们能够提出在蛋白点中每分子蛋白有 1-4 个汞原子的化学计量比。ESI-MS/MS 分析鉴定的蛋白有角蛋白、II 型细胞骨架 8、副肌球蛋白 beta、副肌球蛋白-2、泛素-40S 核糖体 S27a、39S 核糖体蛋白 L36 线粒体、血红蛋白亚基 beta 和血红蛋白亚基 beta-A/B。结果表明,泛素-40S 核糖体 S27a 等具有特定结构域(可能是锌指)的蛋白可能可用作汞的生物标志物,而汞和锌具有软酸的特性。