一种基于活动的综合蛋白质分析方法,用于更高的吞吐量确定蛋白质对小分子的目标敏感性
Chathuri J Kombala1, Agne Sveistyte1, Tong Zhang1
1Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99354, United States.
ACS chemical biology
|August 15, 2025
概括
本研究引入了一种基于活动的综合竞争性蛋白质分析 (ABPP) 方法,以有效地确定蛋白质中化学毒素的敏感性. 新的整体ABPP (IABPP) 方法将样本分析的数量减少了三倍,使得化学品的不良影响可以更快地进行选.
科学领域:
- 化学蛋白质组学 化学蛋白质组学 化学蛋白质组学
- 化学生物学 化学生物学
- 毒理学 毒理学 毒理学
背景情况:
- 基于活动的蛋白质概况 (ABPP) 是一种化学蛋白质学方法,用于标记蛋白质组中的活性酶.
- 竞争性ABPP描述了小分子如何影响蛋白质活动,这对于评估化学物质暴露非常有用.
- 高通量ABPP产生了大量的数据,吸引了受干扰影响的蛋白质标的特征.
研究的目的:
- 开发一个完整的,具有竞争力的ABPP方法来确定化学毒剂的敏感性.
- 在竞争激烈的ABPP中优化协同质量标签 (TMT) 复杂化的条件.
- 为了比较传统的竞争性ABPP与新的聚合样本战略的效率.
主要方法:
- 开发和优化了使用有机酸盐 (OP) 农药作为模型毒剂的综合竞争性ABPP (IABPP) 方法.
- 将传统的竞争性ABPP与各种氧度的集成样本进行比较.
- 验证了使用pyrifos oxon和malaoxon来评估蛋白质标敏感度的方法.
主要成果:
- 聚合竞争样本中的整数强度有效地评估相对蛋白质敏感性,而不会增加样本数量.
- 根据IABPP的策略,对8种化学度的TMT复合和LC-MS/MS分析减少了三倍.
- 鉴定了对各种OP农药的蛋白质点敏感性的差异.
结论:
- 综合ABPP (IABPP) 方法使得对关注化学品 (CoC) 的有效目标灵敏度确定成为可能.
- 这种方法大大减少了查所需的样本和分析数量.
- IABPP提供了一种快速选各种化学物质的手段,以识别高和低敏感性蛋白质标.
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