Department of Bioprocess Engineering, Chonbuk National University, 664-14 Duckjin-dong, Jeonju 561-756, South Korea.
Chemosphere. 2013 Nov;93(10):2341-8. doi: 10.1016/j.chemosphere.2013.08.031. Epub 2013 Sep 12.
In this study, the effects of three widespread heavy metals, As(III), As(V) and Cd, and their binary mixtures on the proteomic profile in D. magna were examined to screen novel protein biomarkers using the two-dimensional gel electrophoresis method (2DE). Ten 20d daphnia were exposed to the LC20 concentrations for each of a total of 8 treatments, including the control, As(III), As(V), Cd, [As(III)+As(V)], [As(III)+Cd], [As(V)+Cd], and [As(III), As(V), Cd], for 24h before protein isolation. Three replicates were performed for each treatment. These protein samples were employed for 2DE experiments with a pH gradient gel strip from pH 3 to pH 10. The protein spots were detected by a silver staining process and their intensities were analyzed by Progenesis software to discover the differentially expressed proteins (DEPs) in response to each heavy metal. A total of 117 differentially expressed proteins (DEPs) were found in daphnia responding to the 8 treatments and mapped onto a 2D proteome map, which provides some information of the molecular weight (MW) and pI value for each protein. All of these DEPs are considered as potential candidates for protein biomarkers in D. magna for detecting heavy metals in the aquatic ecosystem. Comparing the proteomic results among these treatments suggested that exposing D. magna to binary mixtures of heavy metals may result in some complex interactive molecular responses within them, rather than just the simple sum of the proteomic profiles of the individual chemicals, (As(III), As(V), and Cd).
在这项研究中，我们研究了三种广泛存在的重金属（砷(III)、砷(V)和镉）及其二元混合物对大型溞蛋白质组图谱的影响，以使用二维凝胶电泳方法（2DE）筛选新型蛋白质生物标志物。将 10 只 20 天龄的溞暴露于每种 8 种处理的 LC20 浓度下，包括对照、砷(III)、砷(V)、镉、[砷(III)+砷(V)]、[砷(III)+镉]、[砷(V)+镉]和[砷(III)、砷(V)、镉]，持续 24 小时后分离蛋白质。每种处理重复 3 次。这些蛋白质样品用于 2DE 实验，使用 pH 梯度凝胶条从 pH 3 到 pH 10。通过银染过程检测蛋白质斑点，并通过 Progenesis 软件分析其强度，以发现每种重金属处理下差异表达的蛋白质（DEPs）。在溞对 8 种处理的响应中发现了总共 117 个差异表达的蛋白质（DEPs），并映射到 2D 蛋白质组图谱上，该图谱为每种蛋白质提供了分子量（MW）和等电点（pI）值的信息。所有这些 DEPs都被认为是在大型溞中检测水生态系统中重金属的潜在蛋白质生物标志物候选物。比较这些处理之间的蛋白质组学结果表明，暴露于重金属二元混合物中的大型溞可能会导致它们内部产生一些复杂的相互作用分子反应，而不仅仅是个体化学物质（砷(III)、砷(V)和镉）的蛋白质组图谱的简单总和。
