Laboratory for Biological Effects of Metals, Division for Marine and Environmental Research, Ruđer Bošković Institute, P.O. Box 180, 10002 Zagreb, Croatia.
Environ Sci Pollut Res Int. 2013 Apr;20(4):2340-51. doi: 10.1007/s11356-012-1105-8. Epub 2012 Aug 11.
Association of selected essential (Co, Cu, Fe, Mn, Mo, Se, and Zn) and nonessential (Cd, Pb) trace elements with cytosolic proteins of different molecular masses was described for the liver of European chub (Squalius cephalus) from weakly contaminated Sutla River in Croatia. The principal aim was to establish basic trace element distributions among protein fractions characteristic for the fish living in the conditions of low metal exposure in the water. The fractionation of chub hepatic cytosols was carried out by size exclusion high performance liquid chromatography (SE-HPLC; Superdex™ 200 10/300 GL column), and measurements were performed by high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Elution profiles of essential elements were mostly characterized by broad peaks covering wide range of molecular masses, as a sign of incorporation of essential elements in various proteins within hepatic cytosol. Exceptions were Cu and Fe, with elution profiles characterized by sharp, narrow peaks indicating their probable association with specific proteins, metallothionein (MT), and ferritin, respectively. The main feature of the elution profile of nonessential metal Cd was also single sharp, narrow peak, coinciding with MT elution time, and indicating almost complete Cd detoxification by MT under the conditions of weak metal exposure in the water (dissolved Cd concentration ≤0.3 μg L(-1)). Contrary, nonessential metal Pb was observed to bind to wide spectrum of proteins, mostly of medium molecular masses (30-100 kDa), after exposure to dissolved Pb concentration of ~1 μg L(-1). The obtained information within this study presents the starting point for identification and characterization of specific metal/metalloid-binding proteins in chub hepatic cytosol, which could be further used as markers of metal/metalloid exposure or effect on fish.
本文描述了来自克罗地亚弱污染的 Sutla 河的欧洲欧鱈(Squalius cephalus)肝脏中,几种必需(Co、Cu、Fe、Mn、Mo、Se 和 Zn)和非必需(Cd、Pb)微量元素与不同分子量细胞溶质蛋白的关联。其主要目的是确定在水中金属暴露水平较低的情况下,鱼类生存条件下基本微量元素在蛋白质组分中的分布。通过尺寸排阻高效液相色谱法(SE-HPLC;Superdex™ 200 10/300 GL 柱)对欧鱈肝细胞溶质进行分级分离,并用高分辨率电感耦合等离子体质谱法(HR ICP-MS)进行测量。必需元素的洗脱曲线主要表现为覆盖广泛分子量范围的宽峰,表明必需元素在肝细胞溶质中的各种蛋白质中的结合。Cu 和 Fe 是例外,它们的洗脱曲线具有尖锐、狭窄的峰,表明它们可能分别与特定蛋白质(金属硫蛋白 (MT) 和铁蛋白)结合。非必需金属 Cd 的洗脱曲线的主要特征也是单个尖锐、狭窄的峰,与 MT 的洗脱时间一致,表明在水中金属暴露较弱的条件下(溶解 Cd 浓度≤0.3μg L(-1)),几乎完全通过 MT 进行 Cd 解毒。相反,在暴露于约 1μg L(-1)的溶解 Pb 浓度后,非必需金属 Pb 被观察到与广泛的蛋白质结合,主要是中等分子量(30-100 kDa)的蛋白质。本研究中获得的信息为鉴定和表征欧鱈肝细胞溶质中特定金属/类金属结合蛋白提供了起点,这些蛋白可进一步用作金属/类金属暴露或对鱼类影响的标志物。