Quantification of nanocondensates formation at the single molecule level DOI Creative Commons

Justin Houx,

Thomas Copie,

Yann Gambin

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 6, 2024

Abstract Understanding the molecular mechanisms of biomolecular condensate formation through liquid-liquid phase separation is crucial for deciphering cellular cues in normal and pathological contexts. Recent studies have highlighted existence sub-micron assemblies, known as nanocondensates or mesoscopic clusters, organization a significant portion proteome. However, smaller condensates are invisible to classical microscopy, new tools must be developed quantify their numbers properties. Here, we establish simple analysis framework using single molecule fluorescence spectroscopy diffusing solution. We used low-complexity domain TAR DNA-binding protein 43 (TDP-43) model system show that can recapitulate diagram various conditions. Single reveals rapid TDP-43 nanoclusters at ten-fold lower concentrations than described previously by microscopy. demonstrate how straightforward fingerprinting individual provides an exquisite quantification formation, size, density, temporal evolution. Overall, this study highlights potential investigate systems.

Language: Английский

The roles of intrinsically disordered proteins in neurodegeneration DOI Creative Commons
Kagistia Hana Utami, Satoru Morimoto, Yasue Mitsukura

et al.

Biochimica et Biophysica Acta (BBA) - General Subjects, Journal Year: 2025, Volume and Issue: unknown, P. 130772 - 130772

Published: Feb. 1, 2025

Language: Английский

Citations

1
Intrinsic factors behind long COVID: IV. Hypothetical roles of the SARS‐CoV‐2 nucleocapsid protein and its liquid–liquid phase separation DOI

Ahmed Eltayeb,

Faisal Al‐Sarraj, Mona G. Alharbi

et al.

Journal of Cellular Biochemistry, Journal Year: 2024, Volume and Issue: 125(3)

Published: Feb. 13, 2024

Abstract When the SARS‐CoV‐2 virus infects humans, it leads to a condition called COVID‐19 that has wide spectrum of clinical manifestations, from no symptoms acute respiratory distress syndrome. The initiates damage by attaching ACE‐2 protein on surface endothelial cells line blood vessels and using these as hosts for replication. Reactive oxygen species levels are increased during viral replication, which oxidative stress. About three‐fifths (~60%) people who get infected with eradicate their body after 28 days recover normal activity. However, large fraction (~40%) suffer various (anosmia and/or ageusia, fatigue, cough, myalgia, cognitive impairment, insomnia, dyspnea, tachycardia) beyond 12 weeks diagnosed syndrome long COVID. Long‐term studies in group contracted have been contrasted noninfected matched people. A subset can be distinguished set cytokine markers persistent, low‐grade inflammation often self‐report two or more bothersome symptoms. No medication alleviate efficiently. Coronavirus nucleocapsid proteins investigated extensively potential drug targets due key roles among is ability bind respective genomic RNAs incorporation into emerging virions. This review highlights basic its undergo liquid–liquid phase separation. We hypothesize this separation may contribute hypothesis unlocks new investigation angles could potentially open novel avenues better understanding COVID treating condition.

Language: Английский

Citations

6
Membraneless organelles in health and disease: exploring the molecular basis, physiological roles and pathological implications DOI Creative Commons
Yangxin Li, Brian Liu,

Xi‐Yong Yu

et al.

Signal Transduction and Targeted Therapy, Journal Year: 2024, Volume and Issue: 9(1)

Published: Nov. 18, 2024

Abstract Once considered unconventional cellular structures, membraneless organelles (MLOs), substructures involved in biological processes or pathways under physiological conditions, have emerged as central players dynamics and function. MLOs can be formed through liquid-liquid phase separation (LLPS), resulting the creation of condensates. From neurodegenerative disorders, cardiovascular diseases, aging, metabolism to cancer, influence on human health disease extends widely. This review discusses underlying mechanisms LLPS, biophysical properties that drive MLO formation, their implications for We highlight recent advances understanding how physicochemical environment, molecular interactions, post-translational modifications regulate LLPS dynamics. offers an overview discovery current biomolecular condensate conditions diseases. article aims deliver latest insights by analyzing research, highlighting critical role organization. The discussion also covers membrane-associated condensates cell signaling, including those involving T-cell receptors, stress granules linked lysosomes, within Golgi apparatus. Additionally, potential targeting clinical settings is explored, promising avenues future research therapeutic interventions.

Language: Английский

Citations

6
Detection of TurboID fusion proteins by fluorescent streptavidin outcompetes antibody signals and visualises targets not accessible to antibodies DOI Creative Commons
Johanna Odenwald, Bernardo Gabiatti, Silke Braune

et al.

eLife, Journal Year: 2024, Volume and Issue: 13

Published: Aug. 29, 2024

Immunofluorescence localises proteins via fluorophore-labelled antibodies. However, some evade detection due to antibody-accessibility issues or because they are naturally low abundant antigen density is reduced by the imaging method. Here, we show that fusion of target protein biotin ligase TurboID and subsequent biotinylation fluorescent streptavidin offers an ‘all in one’ solution these restrictions. For all tested, signal was significantly stronger than antibody signal, markedly improving sensitivity expansion microscopy correlative light electron microscopy. Importantly, within phase-separated regions, such as central channel nuclear pores, nucleolus, RNA granules, were readily detected with streptavidin, while most antibodies failed. When used tandem HA epitope tag, co-probing anti-HA can map created a for trypanosome pore. Lastly, resolves dynamic, temporally, spatially distinct sub-complexes and, specific cases, reveals history dynamic interaction. In conclusion, has major advantages lowly inaccessible addition, provides information on interactions biophysical environment.

Language: Английский

Citations

5
β-synuclein regulates the phase transitions and amyloid conversion of α-synuclein DOI Creative Commons

Xi Li,

Linwei Yu, Xikai Liu

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Oct. 9, 2024

Parkinson's disease (PD) and Dementia with Lewy Bodies (DLB) are neurodegenerative disorders characterized by the accumulation of α-synuclein aggregates. forms droplets via liquid-liquid phase separation (LLPS), followed liquid-solid (LSPS) to form amyloids, how this process is physiologically-regulated remains unclear. β-synuclein colocalizes in presynaptic terminals. Here, we report that partitions into condensates promotes LLPS, slows down LSPS α-synuclein, while disease-associated mutations lose these capacities. Exogenous improves movement defects prolongs lifespan an α-synuclein-expressing NL5901 Caenorhabditis elegans strain, mutants aggravate symptoms. Decapeptides targeted at α-/β-synuclein interaction sites rationally designed, which suppress rescue defects, prolong C. NL5901. Together, unveil a Yin-Yang balance between α- underlying normal states PD DLB therapeutical potentials.

Language: Английский

Citations

4
The prolyl oligopeptidase and α-synuclein connection revisited DOI
Roos Van Elzen, Yannick Waumans, Sangeeta Nath

et al.

Biochimie, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

Citations

0
Deciphering the role of liquid-liquid phase separation in sarcoma: Implications for pathogenesis and treatment DOI

Zehao Cheng,

Hua Wang, Y M Zhang

et al.

Cancer Letters, Journal Year: 2025, Volume and Issue: 616, P. 217585 - 217585

Published: Feb. 23, 2025

Language: Английский

Citations

0
Probing the Formation and Liquid-to-Solid Transition of FUS Condensates via the Lifetimes of Fluorescent Proteins DOI

Jinyao Ji,

Kui Xu,

Wenjuan Wang

et al.

The Journal of Physical Chemistry Letters, Journal Year: 2025, Volume and Issue: unknown, P. 3553 - 3561

Published: March 31, 2025

Liquid–liquid phase separation (LLPS) of biomolecules is a fundamental cellular process that essential for maintaining homeostasis and facilitating biochemical activities. On the other hand, aberrant alters condensate fluidity causes transition from liquid-like condensates to solid-like condensates, which may lead formation pathological aggregations often observed in neurodegenerative diseases. Condensate usually assessed by fluorescence recovery after photobleaching. Here, we reveal lifetimes several fluorescent proteins are sensitive LLPS liquid-to-solid transition. Furthermore, identify key residues regulate sensitivity toward separation. Thus, apply lifetime imaging microscopy (FLIM) visualize living cells, demonstrating FLIM nondestructive method tracking changes real time.

Language: Английский

Citations

0
Phase separation participates in the genetic regulation mechanism of hematopoietic stem cells: potential therapeutic methods DOI Creative Commons
Xinyu Tang, Yan Wang, Ruirong Xu

et al.

Stem Cell Research & Therapy, Journal Year: 2025, Volume and Issue: 16(1)

Published: May 1, 2025

Language: Английский

Citations

0
Hydrodynamic Radii of Intrinsically Disordered Proteins: Fast Prediction by Minimum Dissipation Approximation and Experimental Validation DOI Creative Commons
Radost Waszkiewicz,

Agnieszka Michaś,

Michał K. Białobrzewski

et al.

The Journal of Physical Chemistry Letters, Journal Year: 2024, Volume and Issue: 15(19), P. 5024 - 5033

Published: May 2, 2024

The diffusion coefficients of globular and fully unfolded proteins can be predicted with high accuracy solely from their mass or chain length. However, this approach fails for intrinsically disordered (IDPs) containing structural domains. We propose a rapid predictive methodology estimating the IDPs. uses accelerated conformational sampling based on self-avoiding random walks includes hydrodynamic interactions between coarse-grained protein subunits, modeled using generalized Rotne−Prager−Yamakawa approximation. To estimate radius, we rely minimum dissipation approximation recently introduced by Cichocki et al. Using large set experimentally measured radii IDPs over wide range lengths domain contributions, demonstrate that our predictions are more accurate than Kirkwood phenomenological approaches. Our technique may prove to valuable in predicting properties both unstructured multidomain proteins.

Language: Английский

Citations

3