Author response: Beyond BioID: Streptavidin outcompetes antibody fluorescence signals in protein localization and readily visualises targets evading immunofluorescence detection DOI Open Access
Johanna Odenwald, Bernardo Gabiatti, Silke Braune

и другие.

Опубликована: Авг. 12, 2024

Immunofluorescence is a common method to localise proteins within their cellular context via fluorophore labelled antibodies and for some applications without alternative. However, protein targets evade detection due low abundance or accessibility issues. In addition, imaging methods require massive reduction in antigen density thus impeding of even medium-abundant proteins.Here, we show that the fusion target TurboID, biotin ligase labelling lysine residues close proximity, subsequent biotinylation by fluorescent streptavidin offers an "all one" solution above-mentioned restrictions. For wide range tested, signal was significantly stronger than antibody signal, markedly improving sensitivity expansion microscopy correlative light electron microscopy, with no loss resolution. Importantly, phase-separated regions, such as central channel nuclear pores, nucleolus RNA granules, were readily detected streptavidin, while most fail label these environments. When TurboID used tandem HA epitope tag, co-probing anti-HA can be map antibody- certain regions. As proof principle, mapped access all trypanosome pore (NUPs) found restricted FG NUPs are known phase-separated, non-FG Nups could labelled. Lastly, resolve dynamic, temporally spatially distinct sub-complexes and, specific cases, reveal history dynamic interaction.In conclusion, has major advantages lowly abundant inaccessible provide information on interactions biophysical environment.

Язык: Английский

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

и другие.

Biochimica et Biophysica Acta (BBA) - General Subjects, Год журнала: 2025, Номер unknown, С. 130772 - 130772

Опубликована: Фев. 1, 2025

Язык: Английский

Процитировано

2

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

Zehao Cheng,

Hua Wang, Y M Zhang

и другие.

Cancer Letters, Год журнала: 2025, Номер 616, С. 217585 - 217585

Опубликована: Фев. 23, 2025

Язык: Английский

Процитировано

2

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

и другие.

Biochimie, Год журнала: 2025, Номер unknown

Опубликована: Фев. 1, 2025

Язык: Английский

Процитировано

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

и другие.

Journal of Cellular Biochemistry, Год журнала: 2024, Номер 125(3)

Опубликована: Фев. 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.

Язык: Английский

Процитировано

9

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

Xi Li,

Linwei Yu, Xikai Liu

и другие.

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Окт. 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.

Язык: Английский

Процитировано

7

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

и другие.

eLife, Год журнала: 2024, Номер 13

Опубликована: Авг. 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.

Язык: Английский

Процитировано

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

и другие.

Signal Transduction and Targeted Therapy, Год журнала: 2024, Номер 9(1)

Опубликована: Ноя. 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.

Язык: Английский

Процитировано

6

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

и другие.

The Journal of Physical Chemistry Letters, Год журнала: 2024, Номер 15(19), С. 5024 - 5033

Опубликована: Май 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.

Язык: Английский

Процитировано

3

Tannic acid as a biphasic modulator of tau protein liquid–liquid phase separation DOI

Jiani Xiang,

Jingxin Chen,

Yanqing Liu

и другие.

International Journal of Biological Macromolecules, Год журнала: 2024, Номер 275, С. 133578 - 133578

Опубликована: Июль 1, 2024

Язык: Английский

Процитировано

3

Serum amyloid A binding to glycosaminoglycans is synergistic with amyloid formation: Therapeutic targeting in the inflammation-linked amyloidosis DOI

Shobini Jayaraman,

Angela Urdaneta,

Marcus Fändrich

и другие.

Journal of Molecular Biology, Год журнала: 2025, Номер unknown, С. 169007 - 169007

Опубликована: Фев. 1, 2025

Язык: Английский

Процитировано

0