Protocols for monitoring condensate formation and dynamics between the phase-separating proteins SET/TAF-Iβ and cytochrome c.

STAR Protoc

Institute for Chemical Research (IIQ), Scientific Research Center "Isla de la Cartuja" (cicCartuja), University of Seville - CSIC, Seville, Spain. Electronic address:

Published: June 2025


Category Ranking

98%

Total Visits

921

Avg Visit Duration

2 minutes

Citations

20

Article Abstract

The targeting of several nuclear stress-response factors by translocated cytochrome c upon genotoxic stress has been demonstrated in recent years to involve liquid-liquid phase separation. This protocol addresses the need to investigate the mechanisms and features of phase separation of the histone chaperone SET/TAF-Iβ induced by cytochrome c. We provide steps for protein purification and fluorescent labeling, condensate formation, imaging, quantification, and evaluation of their dynamics by fluorescence recovery after photobleaching (FRAP). This protocol can be broadly applied to other protein complexes. For complete details on the use and execution of this protocol, please refer to Casado-Combreras et al..

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC12138546PMC
http://dx.doi.org/10.1016/j.xpro.2025.103796DOI Listing

Publication Analysis

Top Keywords

condensate formation
8
phase separation
8
protocols monitoring
4
monitoring condensate
4
formation dynamics
4
dynamics phase-separating
4
phase-separating proteins
4
proteins set/taf-iβ
4
set/taf-iβ cytochrome
4
cytochrome targeting
4

Similar Publications

The solid-solution alloys of Mn-Zn-Ga and Mn-Zn-Sn have been synthesized by a high-temperature method and structurally characterized by X-ray diffraction studies. The substitutional solid-solution alloys that crystallize in the chiral space group 432 or 432 adopt the A13-type structure (β-Mn). Similar to β-Mn, the 20 atoms in the cubic unit cell are distributed over 8 and 12 Wyckoff positions.

View Article and Find Full Text PDF

Nuclear biomolecular condensates are essential sub-compartments within the cell nucleus and play key roles in transcription and RNA processing. Bottom-up construction of nuclear architectures in synthetic settings is non-trivial but vital for understanding the mechanisms of condensates in real cellular systems. Here, we present a facile and versatile synthetic DNA protonucleus (PN) platform that facilitates localized transcription of branched RNA motifs with kissing loops (KLs) for subsequent condensation into complex condensate architectures.

View Article and Find Full Text PDF

Source apportionment of processes contributing to volcanic PM aerosols during the 2021 eruption of Tajogaite.

Sci Total Environ

September 2025

Laboratorio de Materiales para Análisis Químico (MAT4LL), Departamento de Química, Unidad Departamental de Química Analítica, Universidad de La Laguna, Tenerife, Spain.

The 2021-eruption of Tajogaite (La Palma, Canary Islands) was associated with the formation of large amounts of respirable PM aerosols (smaller than 10 μm) that triggered air quality crisis and lockdowns for ∼35,000 persons. This study aims to quantify the contribution of the aerosol formation mechanisms to the volcanic PM concentrations. During the eruption and post-eruption, we monitored trace gases (SO, HF, HCl and NO), and the size distribution and chemical composition of falling-tephra and PM aerosols.

View Article and Find Full Text PDF

CpG-A induces liquid-liquid phase separation of HMGB1 to activate the RAGE-mediated inflammatory pathway.

Proc Natl Acad Sci U S A

September 2025

State Key Laboratory of Green Biomanufacturing, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.

High-mobility group box protein 1 (HMGB1) is a chromatin-associated nonhistone protein widely distributed in the nucleus of eukaryotic cells. It is transported extracellularly as a proinflammatory mediator or late warning protein to induce immune and inflammatory reactions upon stimuli such as microbial infection. Here, we have found that HMGB1 directly interacts with bacterial DNA analogue CpG-A in the extracellular environment to undergo liquid-liquid phase separation (LLPS) via its positively charged DNA-binding domain.

View Article and Find Full Text PDF

This study aimed to synthesize and evaluate the anticancer activity of novel chalcone derivative against colon cancer by in vitro cytotoxicity against HCT-116 (Research Resource Identifiers:CVCL_D4JB) cell line and in vivo using EAC (Research Resource Identifiers: CVCL_1306) and DLA (Research Resource Identifiers: CVCL_VR37) cells inoculated Swiss albino mice. The present study aimed to synthesize the new chalcone derivatives and conduct its anti-colon cancer activity both in vitro and in vivo. The designed compounds were subjected to in silico studies like binding pocket analysis, molecular docking, and ADME studies.

View Article and Find Full Text PDF