通过V型剂进行蛋白质修饰的全面洞察:采用生物对等反应的化学蛋白质学方法
Meining Xing1, Shuo Wang2, Fangfang Cui1
1State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Lifeomics, National Center for Protein Sciences (Beijing), Beijing, China.
Proteomics
|September 1, 2023
概括
有机化合物 (OPs) 通过修改蛋白质而构成威胁. 一个新的生物对等蛋白质组策略有效地识别了人体血中的众多OP蛋白添加物,有助于暴露评估.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 化学生物学 化学生物学
- 毒理学 毒理学 毒理学
背景情况:
- 有机化合物 (OPs) 是有毒剂,可不可逆地改变蛋白质,形成添加物.
- 传统方法很难识别低丰度的OP引证和各种翻译后修改.
研究的目的:
- 在复杂的生物样本中开发一种新的,敏感的蛋白质基因策略来识别有机化合物 (OP) 添加物.
- 在人体血蛋白质组中全面地图化OP加位.
主要方法:
- 采用生物对等的点击-丰富-释放-识别策略,使用基标记的V-代理探针 (AVP) 和生物酸链接器 (BTP-N3).
- 来自人血清白蛋白 (HSA) 或血的被准,通过CuAAC点击化学捕获,并使用斯特列塔维丁珠进行丰富.
- 通过性水解释放了丰富的添加物,并使用液体染色学-并联质谱法 (LC-MS/MS) 识别了它们.
主要成果:
- 该战略确定了人体血中163种蛋白质上的1115个独特的OP添加点.
- 这种方法成功地检测了传统方法遗漏的OP引证.
- 实现了对新型OP基材的全面覆盖.
结论:
- 开发的蛋白质组策略提供了一种敏感和可通用的方法来检测OP-蛋白质添加物.
- 这种方法促进了对毒性OP暴露的后续验证和剂量评估.
- 它扩大了在人类蛋白质组内对OP目标的理解.
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