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作为蛋白质LLPS传感器的ANS染料的时间分辨率光

Sergey A Silonov¹, Semen V Nesterov¹, Anastasia A Gavrilova¹

¹Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., 194064, St. Petersburg, Russia.

Biochemical and biophysical research communications

|December 14, 2024

概括

光染料ANS可以检测水溶液中的内在无序蛋白质中的液态液态相分离 (LLPS). 然而,在巨分子拥挤条件下形成的冷凝物中没有观察到ANS的积累.

科学领域:

生物物理学的生物物理.
分子生物学分子生物学
细胞生物学细胞生物学

背景情况:

内在无序的蛋白质 (IDP) 经历液体-液体相分离 (LLPS),形成生物分子凝聚物.
想象这些凝结物对于理解细胞过程至关重要.
需要新的方法来有效地可视化LLPS.

研究的目的:

调查光染料ANS作为LLPS传感器的实用性.
使用ANS可视化由压力颗粒架构蛋白G3BP1形成的凝聚物.

主要方法:

光终身成像显微镜 (FLIM).
时间分辨率光谱学.
研究了在水溶液和拥挤溶液中的ANS与G3BP1凝结物的相互作用.

主要成果:

在水溶液中的RNA诱导的G3BP1凝结物中积聚的ANS.
在宏分子拥挤 (高PEG度) 下,ANS没有在G3BP1凝聚物中积累.
在G3BP1凝结物的存在下,ANS的限制光异质性减少了.

结论:

ANS显示出作为检测水溶液中的LLPS和IDP的传感器的潜力.
ANS不适合在拥挤条件下形成的G3BP1凝结物的可视化.
时间解析的ANS光可以检测到IDP的LLPS.

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