Language
English
Publication Date
1-1-2023
Journal
Methods in Molecular Biology
DOI
10.1007/978-1-0716-2663-4_6
PMID
36227471
PMCID
PMC10321152
PubMedCentral® Posted Date
7-5-2023
PubMedCentral® Full Text Version
Author MSS
Abstract
Biomolecular condensates of ribonucleoproteins (RNPs) such as the transactivation response element (TAR) DNA-binding protein 43 (TDP-43) arise from liquid-liquid phase separation (LLPS) and play vital roles in various biological processes including the formation-dissolution of stress granules (SGs). These condensates are thought to be directly linked to neurodegenerative diseases, providing a depot of aggregation-prone proteins and serving as a cauldron of protein aggregation and fibrillation. Despite recent research efforts, biochemical processes and rearrangements within biomolecular condensates that trigger subsequent protein misfolding and aggregation remain to be elucidated. Fluorescence lifetime imaging microscopy (FLIM) provides a minimally intrusive high-sensitivity and high-resolution imaging method to monitor in-droplet spatiotemporal changes that initiate and lead to protein aggregation. In this chapter, we describe a FLIM application for characterizing chemical chaperone-assisted decoupling of TDP-43 liquid-liquid phase separation and aggregation/fibrillation, highlighting potential therapeutic strategies to combat pathological RNP-associated aggregates without compromising cellular stress responses.
Keywords
Biomolecular Condensates, DNA-Binding Proteins, Microscopy, Fluorescence, Protein Aggregates, Ribonucleoproteins, Droplet maturation, Fluorescence lifetime imaging microscopy, Intrinsically disordered proteins, Liquid-liquid phase separation, Neurodegenerative diseases, Protein aggregation, TDP-43
Published Open-Access
yes
Recommended Citation
Quan, My Diem; Liao, Shih-Chu Jeff; Ferreon, Josephine C; et al., "Fluorescence Lifetime Imaging Microscopy of Biomolecular Condensates" (2023). Faculty, Staff and Students Publications. 8347.
https://digitalcommons.library.tmc.edu/baylor_docs/8347