利用共聚焦显微镜和光镊测定生物分子凝聚物的热力学和材料性质。
Determining Thermodynamic and Material Properties of Biomolecular Condensates by Confocal Microscopy and Optical Tweezers.
机构信息
Department of Chemistry, University of Illinois at Chicago, Chicago, IL, USA.
Department of Physics, University of Illinois at Chicago, Chicago, IL, USA.
出版信息
Methods Mol Biol. 2023;2563:237-260. doi: 10.1007/978-1-0716-2663-4_12.
Abstract
While the roles of biomolecular condensates in health and disease are being intensely studied, it is equally important that their physical properties are characterized in order to achieve mechanistic understanding. Here we share some of the protocols developed in our lab for measuring thermodynamic and materials properties of condensates. These include a simple method for determining the droplet-phase concentrations of condensate components on a confocal microscope, and a method for determining the viscoelasticity of condensates by optical tweezers. These protocols are either generally applicable to biomolecular condensates or are unique for their characterization.
摘要
虽然生物分子凝聚物在健康和疾病中的作用正在被深入研究，但同样重要的是要对其物理性质进行特征描述，以实现对其机制的理解。在这里，我们分享了一些在我们实验室中开发的用于测量凝聚物热力学和材料性质的方案。这些方案包括在共聚焦显微镜上确定凝聚物组分液滴相浓度的简单方法，以及通过光镊确定凝聚物粘弹性的方法。这些方案要么普遍适用于生物分子凝聚物，要么是其特征描述所特有的。
相似文献
1
Determining Thermodynamic and Material Properties of Biomolecular Condensates by Confocal Microscopy and Optical Tweezers.利用共聚焦显微镜和光镊测定生物分子凝聚物的热力学和材料性质。
Methods Mol Biol. 2023;2563:237-260. doi: 10.1007/978-1-0716-2663-4_12.
2
Programmable viscoelasticity in protein-RNA condensates with disordered sticker-spacer polypeptides.具有无序黏附-间隔多肽的蛋白质-RNA 凝聚物中的可编程粘弹性。
Nat Commun. 2021 Nov 16;12(1):6620. doi: 10.1038/s41467-021-26733-7.
3
Methods for characterizing the material properties of biomolecular condensates.表征生物分子凝聚物材料特性的方法。
Methods Enzymol. 2021;646:143-183. doi: 10.1016/bs.mie.2020.06.009. Epub 2020 Jul 22.
4
Time-Dependent Material Properties of Aging Biomolecular Condensates from Different Viscoelasticity Measurements in Molecular Dynamics Simulations.基于分子动力学模拟的不同粘弹性测量的老化生物分子凝聚物的时变材料特性。
J Phys Chem B. 2023 May 25;127(20):4441-4459. doi: 10.1021/acs.jpcb.3c01292. Epub 2023 May 17.
5
Shape recovery of deformed biomolecular droplets: Dependence on condensate viscoelasticity.变形生物分子液滴的形状恢复：对凝聚态粘弹性的依赖。
J Chem Phys. 2021 Oct 14;155(14):145102. doi: 10.1063/5.0064247.
6
Shear relaxation governs fusion dynamics of biomolecular condensates.剪切松弛控制生物分子凝聚体的融合动力学。
Nat Commun. 2021 Oct 13;12(1):5995. doi: 10.1038/s41467-021-26274-z.
7
Label-Free Techniques for Probing Biomolecular Condensates.无标记技术在探测生物分子凝聚物中的应用。
ACS Nano. 2024 Apr 23;18(16):10738-10757. doi: 10.1021/acsnano.4c01534. Epub 2024 Apr 12.
8
Macromolecular Regulation of the Material Properties of Biomolecular Condensates.生物分子凝聚物材料特性的大分子调控
J Phys Chem Lett. 2022 Jun 8:5285-5290. doi: 10.1021/acs.jpclett.2c00824.
9
Macromolecular regulators have matching effects on the phase equilibrium and interfacial tension of biomolecular condensates.大分子调节剂对生物分子凝聚相的相平衡和界面张力具有匹配的影响。
Protein Sci. 2021 Jul;30(7):1360-1370. doi: 10.1002/pro.4084. Epub 2021 Apr 24.
10
Surface tension measurement and calculation of model biomolecular condensates.表面张力测量和模型生物分子凝聚体的计算。
Soft Matter. 2023 Nov 22;19(45):8706-8716. doi: 10.1039/d3sm00820g.
引用本文的文献
1
Homotypic RNA clustering accompanies a liquid-to-solid transition inside the core of multi-component biomolecular condensates.同型RNA聚类伴随着多组分生物分子凝聚物核心内的液-固转变。
Nat Chem. 2025 Jul 2. doi: 10.1038/s41557-025-01847-3.
2
Amino acid-dependent phase equilibrium and material properties of tetrapeptide condensates.四肽缩合物的氨基酸依赖性相平衡和材料特性
Cell Rep Phys Sci. 2024 Oct 16;5(10). doi: 10.1016/j.xcrp.2024.102218. Epub 2024 Sep 23.
3
Biomolecular condensates can enhance pathological RNA clustering.生物分子凝聚物可增强病理性RNA聚集。
Res Sq. 2024 Jul 19:rs.3.rs-4557520. doi: 10.21203/rs.3.rs-4557520/v1.
4
Biomolecular Condensates Can Enhance Homotypic RNA Clustering.生物分子凝聚物可增强同型RNA聚集。
bioRxiv. 2025 Jan 20:2024.06.11.598371. doi: 10.1101/2024.06.11.598371.
5
Amino Acid-Dependent Material Properties of Tetrapeptide Condensates.四肽缩合物的氨基酸依赖性材料特性
bioRxiv. 2024 May 15:2024.05.14.594233. doi: 10.1101/2024.05.14.594233.
6
Adenosine Triphosphate Mediates Phase Separation of Disordered Basic Proteins by Bridging Intermolecular Interaction Networks.三磷酸腺苷通过桥接分子间相互作用网络介导无序碱性蛋白的液-液相分离。
J Am Chem Soc. 2024 Jan 17;146(2):1326-1336. doi: 10.1021/jacs.3c09134. Epub 2024 Jan 4.
7
A calibration-free model of micropipette aspiration for measuring properties of protein condensates.无校准微管吸吮模型用于测量蛋白质凝聚物的性质。
Biophys J. 2024 Jun 4;123(11):1393-1403. doi: 10.1016/j.bpj.2023.09.018. Epub 2023 Oct 2.
8
ATP Mediates Phase Separation of Disordered Basic Proteins by Bridging Intermolecular Interaction Networks.ATP通过桥接分子间相互作用网络介导无序碱性蛋白的相分离。
bioRxiv. 2023 Aug 20:2023.08.20.554035. doi: 10.1101/2023.08.20.554035.
本文引用的文献
1
Surface tension and viscosity of protein condensates quantified by micropipette aspiration.通过微量移液器抽吸法定量蛋白质凝聚物的表面张力和粘度。
Biophys Rep (N Y). 2021 Sep 8;1(1). doi: 10.1016/j.bpr.2021.100011. Epub 2021 Aug 11.
2
Programmable viscoelasticity in protein-RNA condensates with disordered sticker-spacer polypeptides.具有无序黏附-间隔多肽的蛋白质-RNA 凝聚物中的可编程粘弹性。
Nat Commun. 2021 Nov 16;12(1):6620. doi: 10.1038/s41467-021-26733-7.
3
Shear relaxation governs fusion dynamics of biomolecular condensates.剪切松弛控制生物分子凝聚体的融合动力学。
Nat Commun. 2021 Oct 13;12(1):5995. doi: 10.1038/s41467-021-26274-z.
4
Quantitative theory for the diffusive dynamics of liquid condensates.液体凝聚态扩散动力学的定量理论。
Elife. 2021 Oct 12;10:e68620. doi: 10.7554/eLife.68620.
5
Viscoelasticity of biomolecular condensates conforms to the Jeffreys model.生物分子凝聚物的粘弹性符合杰弗里斯模型。
J Chem Phys. 2021 Jan 28;154(4):041103. doi: 10.1063/5.0038916.
6
Surface tensiometry of phase separated protein and polymer droplets by the sessile drop method.固着液滴法研究相分离蛋白和聚合物液滴的表面张力。
Soft Matter. 2021 Feb 19;17(6):1655-1662. doi: 10.1039/d0sm01319f.
7
Protein condensates as aging Maxwell fluids.蛋白质凝聚物作为老化的麦克斯韦流体。
Science. 2020 Dec 11;370(6522):1317-1323. doi: 10.1126/science.aaw4951.
8
Determination of Condensate Material Properties from Droplet Deformation.通过液滴变形确定冷凝物材料特性
J Phys Chem B. 2020 Sep 24;124(38):8372-8379. doi: 10.1021/acs.jpcb.0c06230. Epub 2020 Sep 16.
9
Determinants for Fusion Speed of Biomolecular Droplets.生物分子液滴融合速度的决定因素。
Angew Chem Int Ed Engl. 2020 Nov 16;59(47):20837-20840. doi: 10.1002/anie.202006711. Epub 2020 Sep 8.
10
Tug of War between Condensate Phases in a Minimal Macromolecular System.在最小的高分子体系中凝聚相之间的拔河比赛。
J Am Chem Soc. 2020 May 13;142(19):8848-8861. doi: 10.1021/jacs.0c01881. Epub 2020 May 4.
Zotero 插件