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用光学镊子评估染色质结合酶对浓缩染色质的影响。

Assessing the Effect of Chromatin-Binding Enzymes on Condensed Chromatin with Optical Tweezers.

作者信息

Kamp Dieter, Vizjak Petra, Stigler Johannes

机构信息

Gene Center, Department of Biochemistry, Ludwig-Maximilians-Universität München, Munich, Germany.

Institute of Physiological Chemistry, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.

出版信息

Methods Mol Biol. 2025;2881:345-355. doi: 10.1007/978-1-0716-4280-1_17.

Abstract

The formation of biomolecular condensates in vitro and in vivo has become an increasingly important subject of studies. One particular area of interest is the phase separation of chromatin in the nucleus. However, the interplay of condensed chromatin and chromatin-binding enzymes has barely been studied as of now. Here, we show an optical tweezer-based assay that uses controlled fusion of two condensates to probe the effect of enzymes, such as chromatin remodeling motor proteins, on the fluidity of condensates. The assay provides a powerful tool that enables the study of if and how chromatin-enzyme interactions alter biophysical properties in these dense chromatin condensates.

摘要

体外和体内生物分子凝聚物的形成已成为一个越来越重要的研究课题。一个特别受关注的领域是细胞核中染色质的相分离。然而,到目前为止,凝聚染色质与染色质结合酶之间的相互作用几乎尚未得到研究。在此,我们展示了一种基于光镊的检测方法,该方法利用两个凝聚物的可控融合来探究诸如染色质重塑运动蛋白等酶对凝聚物流动性的影响。该检测方法提供了一个强大的工具,能够研究染色质 - 酶相互作用是否以及如何改变这些致密染色质凝聚物的生物物理性质。

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