使用AirID结合的抗原结合片段对EGFR进行细胞外蛋白与蛋白相互作用的近距离分析
Kohdai Yamada1, Ryouhei Shioya1, Kohei Nishino2
1Division of Cell-Free Life Science, Proteo-Science Center, Ehime University, 3 Bunkyo-cho, Matsuyama, Ehime, 790-8577, Japan.
Nature communications
|December 14, 2023
概括
我们使用FabID开发了一种新方法来分析表皮生长因子受体 (EGFR) 的细胞外蛋白相互作用. 这种技术成功地识别了已知的和新的EGFR相互作用体,显示了对连接体和药物治疗的反应.
科学领域:
- 细胞生物学 细胞生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 像表皮生长因子受体 (EGFR) 这样的受体蛋白通过细胞外蛋白与蛋白相互作用 (exPPI) 中介关键的细胞内信号传递.
- 了解exPPI对于阐明细胞表面受体的生物功能至关重要.
研究的目的:
- 开发和验证一种新的靠近依赖生物化方法,用于分析膜受体蛋白的exPPIs.
- 研究EGFR的exPPI场景及其通过连接体结合和激酶抑制的调制.
主要方法:
- 设计了一种融合蛋白,EGFR-FabID,将生物化酶 (AirID) 与EGFR特异性抗体片段 (Fab) 结合起来.
- 利用EGFR-FabID用于各种细胞系中EGFR细胞外域的靠近依赖生物化.
- 使用液体染色学-质谱学 (LC-MS/MS) 来识别代表 exPPI 的生物化蛋白质.
主要成果:
- 成功确定了许多已知的EGFR相互作用体和参与exPPI的新型候选蛋白.
- 证明已识别的exPPIs是通过表皮生长因子 (EGF) 连接体刺激动态调节的.
- 在使用EGFR激酶抑制剂gefitinib治疗后观察到exPPI的显著变化.
结论:
- 基于FabID的近距离标签是一种有效的策略,用于对细胞表面受体进行公正的exPPI分析.
- 这种方法提供了对EGFR介导的信号通路及其通过外部刺激和药理学剂的调节的洞察.
- 这些发现凸显了FabID在药物发现和理解受体-连接体-药物相互作用方面的实用性.
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