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在单个活细胞中,CRDBP蛋白相分离及其对β-catenin蛋白的招募
Jian Liu1, Shengrong Yu1, Wenxin Yu1
1School of Chemistry & Chemical Engineering, State Key Laboratory of Metal Matrix Composites, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, P. R. China.
The journal of physical chemistry. B
|December 5, 2023
概括
我们开发了一种现场方法来研究活细胞中的编码区域决定性结合蛋白 (CRDBP) 阶段分离. 这项技术观察了CRDBP在单体,集群和颗粒状态,揭示了它在细胞过程中的作用.
科学领域:
- 生物化学 生化学
- 细胞生物学 细胞生物学
- 生物物理学的生物物理.
背景情况:
- 编码区决定性结合蛋白 (CRDBP) 在癌细胞中过度表达,形成颗粒.
- 在CRDBP中,内在无序区域 (IDR) 暗示了潜在的液态-液态相分离 (LLPS).
- 缺乏体内方法阻碍了对细胞内蛋白相分离的研究.
研究的目的:
- 开发一种有效的现场方法,用于研究活细胞中CRDBP相位分离.
- 为了在体内研究CRDBP的相分离行为.
- 探索CRDBP结构和抑制剂对其相位分离和招募的影响.
主要方法:
- 在现场光相关谱 (FCS) 和光交叉相关谱 (FCCS) 与光蛋白融合相结合.
- 在溶液中使用光成像和FCS预测和确认了CRDBP相分离.
- 研究了CRDBP相分离状态 (单体,集群,颗粒) 和活细胞中的抑制作用.
主要成果:
- 成功建立了一个现场方法来观察单个活细胞中的CRDBP相位分离.
- 确定了三种不同的CRDBP相分离状态:单体,集群和颗粒.
- 研究了CRDBP截断形式和抑制剂对相分离和β-catenin招募的影响.
- 发现CRDBP对β-catenin的招募,可能与Wnt信号进行链接.
结论:
- FCS/FCCS为研究细胞内蛋白相分离提供了一种高效且替代的现场方法.
- 了解CRDBP阶段分离提供了有关癌症机制和Wnt信号调节的见解.
- 开发的方法使得蛋白质相分离动态的体内研究成为可能.
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