通过生物发光触发的光催化近距离标记,通过精确的和在生物体中兼容的空间蛋白质组学
Xuege Sun1,2, Yanling Zhang1, Wenjie Lu1
1School of Pharmaceutical Sciences, Tsinghua University, Beijing 100084, China.
ACS central science
|September 29, 2025
概括
基于生物发光共振能量转移的识别 (BRET-ID) 在体内精确地绘制了蛋白质相互作用和器官的定位. 这种新的近距离标记技术为动态生物过程提供了高空间和时间分辨率.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 蛋白质局部化和相互作用对于细胞功能至关重要.
- 传统的近距离标签 (PL) 方法面临毒性,高背景和受限制的体内应用等局限性.
- 可见光触发的PL受光诱导的背景和有限的体内使用所阻碍.
研究的目的:
- 开发一种与体内相容的近距离标记技术,用于精确地绘制无膜有机体和短暂的蛋白质与蛋白质相互作用的地图.
- 为了研究动态生物事件,实现分分钟时间分辨率.
- 为了克服现有的PL方法的局限性.
主要方法:
- 开发BRET-ID,结合一个基因编码的光催化剂和NanoLuc luciferase.
- 利用生物发光共振能量转移 (BRET) 在本地产生蓝光并激活光催化剂.
- 采用单片氧来氧化蛋白质,并通过精简的化学蛋白质组工作流程进行后续分析.
主要成果:
- 通过BRET-ID,可以精确地绘制具有高空间特异性的ER膜蛋白.
- 该技术提供了1分钟的动态GPCR相互作用的快照在联体诱导内细胞分裂期间.
- 在压力细胞和瘤异种移植中,BRET-ID确定了新的压力颗粒成分,包括RICTOR.
结论:
- BRET-ID是一种强大的,基因编码的工具,用于体内近距离标签.
- 它允许精确地绘制蛋白质定位和高空间和时间分辨率的分子相互作用.
- BRET-ID促进了动态细胞过程的研究和在生物体中发现新型蛋白相互作用.
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