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

砷结合蛋白的表征及其在健康效果评价中的应用。

• 批准号: 23390167
• 负责人: HIRANO Seishiro
• 金额: $ 11.23万
• 依托单位: National Institute for Environmental Studies
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23390167/
• 关键词: 環境中毒; ヒ素; タンパク質; 電気泳動; promyelocytic leukemia; SUMO化; 蛍光染色; キャラクタリゼーション; 遺伝子導入; 細胞; 毒性; Promyelocytic leukemia; 結合タンパク質; 担体

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.

我们制备了砷结合的琼脂糖凝胶，并通过色谱法使用填充有砷结合的琼脂糖凝胶的柱来研究砷结合蛋白。SDS-PAGE分析表明，没有特异性结合五价砷的蛋白质，而几个蛋白质具有特异性和高亲和力的三价砷。 我们制备了表达早幼粒细胞白血病（PML）的细胞，并将胞浆蛋白施加到砷结合柱上，用含谷胱甘肽的溶液洗脱柱。 PML似乎通过在该蛋白的RING指结构域形成三元复合物来结合三价砷。免疫印迹和免疫荧光显微镜分析表明，PML与小泛素样修饰蛋白（SUMO化）结合，以响应砷暴露。 砷结合的琼脂糖凝胶柱使我们能够确定蛋白质具有高亲和力的砷使用这种方法。

项目成果

多剤耐性タンパク質MRP2 高発現細胞における細胞内ヒ素代謝・排泄機構の解析

高表达多药耐药蛋白MRP2细胞内砷代谢及排泄机制分析

• 发表时间: 2012
• 作者: 渡辺喬之;小林弥生;平野靖史郎
• 通讯作者: 平野靖史郎

Western blot analysis. Methods Mol Biol, In Nonotoxicology : Methods and Protocol

蛋白质印迹分析。

• 发表时间: 2012
• 作者: Hirano;Joshua L. Reineke
• 通讯作者: Joshua L. Reineke

The role of glutathione in the metabolism of diphenylarsinic acid in rats

• DOI: 10.1039/c2mt20228j
• 发表时间: 2013-01-01
• 期刊: METALLOMICS
• 影响因子: 3.4
• 作者: Kobayashi, Yayoi;Hirano, Seishiro
• 通讯作者: Hirano, Seishiro

Developmental subchronic exposure to diphenylarsinic acid induced increased exploratory behavior, impaired learning behavior, and decreased cerebellar glutathione concentration in rats.

• DOI: 10.1093/toxsci/kft200
• 发表时间: 2013-12
• 期刊: Toxicological sciences : an official journal of the Society of Toxicology
• 作者: Negishi T;Matsunaga Y;Kobayashi Y;Hirano S;Tashiro T
• 通讯作者: Tashiro T

多剤耐性タンパク質MRP2高発現細胞における細胞内ヒ素代謝・排泄機構の解析

高表达多药耐药蛋白MRP2细胞内砷代谢及排泄机制分析

• 发表时间: 2012
• 作者: 渡辺喬之;小林弥生;平野靖史郎
• 通讯作者: 平野靖史郎