蛋白质中小分子结合部位的选和识别
Jun Ando1,2,3,4, Miwako Asanuma1,2,3, Kosuke Dodo1,2,3
1AMED-CREST, Japan Agency for Medical Research and Development , Saitama 351-0198, Japan.
Journal of the American Chemical Society
|October 11, 2016
概括
阿尔基因标签拉曼选 (ATRaS) 识别了蛋白质上的小分子结合点. 这种敏感的方法使用拉曼散射和质谱来发现药物和蛋白质组.
科学领域:
- 化学生物学
- 蛋白质组学
- 药物发现
背景情况:
- 识别蛋白质上的小分子结合点对于药物发现和蛋白质功能的理解至关重要.
- 目前使用液态染色体质谱法 (LC-MS) 的方法通常受到复杂混合物的限制.
- 可以识别修饰的氨基酸, 但效率是一个挑战.
研究的目的:
- 开发一种新的,高度敏感的方法来识别蛋白质上的小分子结合点.
- 克服分析复杂混合物的现有技术的局限性.
- 提高药物发现和蛋白质组学的LC-MS分析的选择性和效率.
主要方法:
- 使用与蛋白质形成共价键的标小分子开发了标拉曼选 (ATRaS).
- 使用表面增强的拉曼散射 (SERS) 用银纳米粒子在蛋白质分解和HPLC后敏感检测标记.
- 使用MS/MS来确定结合位置.
主要成果:
- 在选中,ATRaS表现出高灵敏度 (∼100 femtomole检测极限) 和可重复性.
- 通过自动化ATRaS系统成功确定了囊蛋白酶甲素B的抑制剂结合部位.
- 用复杂的混合物验证ATRaS系统的有效性,例如素消化细胞溶解物.
结论:
- ATRaS技术提供高分子选择性,显著改善LC-MS分析.
- 该方法在药物发现,蛋白质组学,代谢学和化学生物学方面具有广泛的潜在应用.
- ATRaS提供了一种灵敏且可重复的方法来识别蛋白质 - 连接体相互作用.
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