Specific capture of uranyl protein targets by metal affinity chromatography
Specific capture of uranyl protein targets by metal affinity chromatography
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DOI：
10.1016/j.chroma.2008.01.081
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发表时间：
2008-03-28
影响因子：
4.1
通讯作者：
Vidaud, Claude
中科院分区：
文献类型：
--
作者：
Basset, Christian;Dedieu, Alain;Vidaud, Claude
为了提高对铀毒理学领域生化机制的总体了解，需要加强对蛋白质靶标的识别。固定化金属亲和层析(IMAC)作为一种从生物提取物中捕获金属结合蛋白的有力工具而得到广泛发展。然而，铀酰离子(UO22+)具有特殊的物理化学性质，使其不能固定在经典的金属络合载体上。本文首次报道了一种基于氨基膦酸基与铀酰结合的阳离子交换性质的固定化铀酰亲和层析方法。测定了阳离子在载体上的分配系数和负载能力。然后在我们的层析条件下对铀酰键合相的稳定性进行了优化，以促进亲和机理。然后通过蛋白质组学和质谱学分析，证明成功地从人血清中富集了铀酰结合蛋白。(C)2008爱思唯尔B.V.保留所有权利。
To improve general understanding of biochemical mechanisms in the field of uranium toxicology, the identification of protein targets needs to be intensified. Immobilized metal affinity chromatography (IMAC) has been widely developed as a powerful tool for capturing metal binding proteins from biological extracts. However uranyl cations (UO22+) have particular physico-chemical characteristics which prevent them from being immobilized on classical metal chelating supports. We report here on the first development of an immobilized uranyl affinity chromatography method, based on the cation-exchange properties of aminophosphonate groups for uranyl binding. The cation distribution coefficient and loading capacity on the support were determined. Then the stability of the uranyl-bonded phase under our chromatographic conditions was optimized to promote affinity mechanisms. The successful enrichment of uranyl binding proteins from human serum was then proven using proteomic and mass spectral analysis. (C) 2008 Elsevier B.V. All rights reserved.