Characterization of arsenic-binding proteins and its application to health effect evaluation.

我们制备了砷结合的sepharose，并利用挤满砷结合sepharose的色谱柱对砷结合蛋白进行了研究。SDS-PAGE分析显示，没有蛋白质与五价砷特异性结合，而一些蛋白质对三价砷具有特异性和高亲和力。制备早幼粒细胞白血病（PML）表达细胞，将胞质蛋白应用于砷结合柱，用含谷胱甘肽溶液洗脱。PML似乎通过在该蛋白环指结构域形成三元配合物与三价砷结合。免疫印迹和免疫荧光显微镜分析表明，砷暴露后PML与小泛素样修饰蛋白结合（SUMOylation）。砷结合的sepharose柱使我们能够用这种方法鉴定对砷有高亲和力的蛋白质。

We prepared arsenic-bound sepharoses and investigated arsenic-binding proteins by chromatography using a column packed with the arsenic-bound sepharoses. SDS-PAGE analyses showed no proteins bound to pentavalent arsenic specifically, while several proteins that have specific and high affinity for trivalent arsenic. We prepared promyelocytic leukemia (PML) expressing cells and applied the cytosolic proteins to the arsenic-binding column and the column was elutes with glutathione-containing solution. PML appeared to bind trivalent arsenic by forming ternary complex at RING finger domain of this protein. Western blot and immunofluorescent microscopic analyses suggested that PML were conjugation with small ubiquitin-like modifier proteins (SUMOylation) in response to arsenic exposure. The arsenic-bound sepharose column enabled us to identify proteins which have a high affinity to arsenic using this method.