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用染色学指导的同分离质谱技术快速分析受药物干扰的蛋白质复合物

Wei Liu¹, Jiayu Tang², Guoquan Yan²

¹Center for Medical Research and Innovation, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 201399, China.

Analytical chemistry

|August 29, 2025

概括

我们开发了一种快速的方法,XL-CoFrac-Q-MS, 这种技术精确量化了蛋白质复杂的变化,有助于选药物标并了解细胞反应.

科学领域:

蛋白质组学
化学生物学
分子细胞生物学

背景情况:

蛋白质复合体对于细胞功能和药物反应至关重要.
分析蛋白质复合物的传统联合分离质谱法 (CoFrac-MS) 是缓慢而劳动密集的.
药物发现需要快速识别受药物干扰的蛋白质复合体.

研究的目的:

引入和验证一种新的,以快速染色学为指导的策略,用于识别受药物干扰的蛋白质复合体.
建立XL-CoFrac-Q-MS工作流程,用于高分辨率分离,精确量化和蛋白质复合物的差异分析.
证明工作流在识别药物向相互作用和阐明药物机制方面的实用性.

主要方法:

开发了XL-CoFrac-Q-MS工作流程,将交叉链接增强的逆相液体染色学联合分离 (XL-CoFrac) 与ChromaQuant软件进行峰值量化.
使用ChromaQuant进行精确的峰值量化 (CV<1%) 和蛋白质复合物的差异分析.
应用工作流来分析用BCL-2抑制剂治疗的白血病细胞的药物反应.

主要成果:

通过XL-CoFrac-Q-MS可快速精确地对蛋白质复合体进行分析,其可重复性很高 (R > 0. 98).
工作流程成功识别了药物扰乱的蛋白质复合体,包括与BCL-2相关的复合体,作为对ABT-199治疗的反应.
鉴定了BCL-2和FKBP38之间的交联,可能揭示药物耐药性的机制.
路径丰富分析提供了关于ABT-199诱导的亡的见解.

结论:

XL-CoFrac-Q-MS策略为快速识别受药物干扰的蛋白质复合物提供了显著的进步.
这种工作流程促进了有效的药物选和在药理学和病理学上阐明蛋白质复杂的动态.
这种方法对了解细胞对各种干扰的反应具有广泛的潜力.

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