文章基本信息

标题：A light-switching pyrene probe to detect phase-separated biomolecules
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作者：Masaharu Hazawa ; Shogo Amemori ; Yoshio Nishiyama 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2021
卷号：24
期号：8
页码：1-15
DOI：10.1016/j.isci.2021.102865
语种：English
出版社：Elsevier
摘要：SummaryBiomolecules may undergo liquid–liquid phase separation (LLPS) to spatiotemporally compartmentalize and regulate diverse biological processes. Because the number of tools to directly probe LLPS is limited (ie. FRAP, FRET, fluorescence microscopy, fluorescence anisotropy, circular dichroism, etc.), the physicochemical traits of phase-separated condensates remain largely elusive. Here, we introduce a light-switching dipyrene probe (Pyr-A) that forms monomers in either hydrophobic or viscous environments, and intramolecular excimers in aqueous solutions. By exploiting their distinct fluorescence emission spectra, we used fluorescent microscopic imaging to study phase-separated condensates formed byin vitroprotein droplets and membraneless intracellular organelles (centrosomes). Ratiometric measurement of excimer and monomer fluorescence intensities showed that protein droplets became hydrophobic and viscous as their size increased. Moreover, centrosomes became hydrophobic and viscous during maturation. Our results show that Pyr-A is a valuable tool to characterize LLPS and enhance our understanding of phase separation underlying biological functions.Graphical abstractDisplay OmittedHighlights•A dipyrene probe Pyr-A is able to monitor LLPS without complicated steps•Pyr-A switches excimers (E) to monomers (M) when environments became hydrophobic/viscous•Ratio-metric measurement (E/M) can evaluate physicochemical dynamics of LLPS•Phase-separated biomolecules became hydrophobic and viscous during increase in sizeBiophysical chemistry; Biological sciences; Biochemistry; Biophysics; Biological sciences research methodologies