原子分辨率相互作用调节FUS折叠RRM域的分离成模型CAPRIN1凝聚物
Rashik Ahmed1,2,3,4, Jeffrey P Bonin1,2,3, Julie D Forman-Kay3,4
1Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.
Journal of the American Chemical Society
|August 26, 2025
概括
这项研究揭示了折叠FUSRNA识别因子 (RRM) 蛋白如何与生物分子凝聚物中的CAPRIN1支架蛋白结合. 这些相互作用驱动选择性客户端丰富,并被CAPRIN1酸化破坏.
科学领域:
- 生物化学
- 分子生物学
- 结构生物学
背景情况:
- 生物分子凝聚剂通过选择性缩分子来调节细胞功能.
- 了解客户端分区需要研究与折叠和展开的客户端状态的相互作用.
- FUS RNA识别因子 (RRM) 是一个客户端蛋白质,而CAPRIN1 是一个参与凝结物形成的支架蛋白.
研究的目的:
- 在相分离过程中研究折叠FUS RRM和CAPRIN1之间的原子级相互作用.
- 确定这些相互作用如何调解CAPRIN1凝聚物中的FUS RRM的选择性丰富.
- 探索 CAPRIN1 后翻译修改对客户端分区的影响.
主要方法:
- 溶液核磁共振 (NMR) 光谱,包括 [1H-15N]-HSQC,分子间核超振效应 (NOE) 和偏磁放松增强 (PRE).
- 化学转移扰动分析以绘制相互作用地点.
- 在40°C的CAPRIN1冷凝液中研究FUS RRM分离.
主要成果:
- 在40°C时,约40%的FUS RRM仍然被折叠在CAPRIN1凝聚物中,允许进行高分辨率的NMR研究.
- 在FUS RRM上确定了与CAPRIN1的芳香和氨酸丰富区域结合的特定相互作用表面.
- 在凝聚物中观察到FUS RRM的30倍丰富,在架-架和架-客户端相互作用中重叠结合点.
- CAPRIN1的氨酸酸化显著破坏了FUS RRM的结合,并减少了超过100倍的分离.
结论:
- 折叠FUS RRM与CAPRIN1进行特定的异型相互作用,导致其选择性分割为生物分子凝聚物.
- 脚手架-客户端和脚手架-脚手架识别具有共同的交互接口,这表明了凝聚剂组装的统一机制.
- 转化后的修改,如CAPRIN1的氨酸酸化,通过调节客户互动来作为凝结物组成的关键调节剂.
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