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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Updated: Jul 5, 2025

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

Published on: December 1, 2018

在液态-液态相分离过程中检测蛋白质-蛋白质相互作用,使用化蛋白质传感器.

Yanan Huang¹, Junlin Chen¹, Chia-Heng Hsiung¹

¹Department of Chemistry, Research Center for Industries of the Future, Westlake University, Hangzhou 310030, Zhejiang, China.

Molecular biology of the cell

|January 17, 2024

概括

分化依赖光蛋白 (ddFPs) 能够在液体-液体相分离 (LLPS) 过程中检测蛋白相互作用. 这种化学生物学方法揭示了RNARNA.

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A Fluorescence Fluctuation Spectroscopy Assay of Protein-Protein Interactions at Cell-Cell Contacts

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科学领域:

生物化学生物化学
细胞生物学细胞生物学
分子生物学分子生物学

背景情况:

细胞凝结物,类似于无膜有机体,通过液体-液体相分离 (LLPS) 形成.
了解蛋白质和RNA相互作用对于阐明LLPS机制和相关疾病至关重要.
由于衍射极限,传统的光显微镜难以可靠地检测蛋白质相互作用.

研究的目的:

开发一种新的化学生物学方法,用于在LLPS过程中检测蛋白质相互作用.
研究RNA在凝聚物中RNA结合蛋白 (RBPs) 之间的相互作用中介作用的作用.

主要方法:

作为一种新的化学生物学方法,利用了依赖于二分化的光蛋白 (ddFPs).
在实验室和活细胞中,在LLPS期间应用ddFP来检测蛋白质与蛋白质相互作用.
研究了FUS和TDP-43之间的相互作用,这两种关键蛋白涉及LLPS.

主要成果:

在LLPS期间成功使用ddFP来可视化和确认蛋白质相互作用.
证明了FUS和TDP-43在形成细胞凝结物中的相互作用.
发现FUS和TDP-43之间的相互作用与LLPS期间的RNA结合有关.

结论:

ddFPs提供了一种可靠的方法来检测LLPS中的蛋白相互作用.
在凝结物形成过程中,RNA在RBPs之间的相互作用中起着关键作用.
ddFPs具有广泛的潜力,可以揭示新型蛋白相互作用及其在生物分子凝聚物动态中的功能.