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.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Дек. 2, 2023

ABSTRACT 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. 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

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

8

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

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

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

The Regulation of TDP-43 Structure and Phase Transitions: A Review DOI

Yanqing Liu,

Jiani Xiang,

Hang Gong

и другие.

The Protein Journal, Год журнала: 2025, Номер unknown

Опубликована: Фев. 22, 2025

Язык: Английский

Процитировано

0

Caffeine Inhibits Tau Aggregation and Destabilizes the Fibril Associated with Chronic Traumatic Encephalopathy: A REST2 and Conventional MD Simulations Study DOI
Jiaxing Tang, Zhengdong Xu, Feng Wang

и другие.

Journal of Chemical Information and Modeling, Год журнала: 2025, Номер unknown

Опубликована: Март 7, 2025

Chronic traumatic encephalopathy (CTE) is a unique tauopathy mostly diagnosed in contact sports athletes, such as those active American football, boxing, soccer, etc. The hyperphosphorylated fibrillar aggregates composed of self-assembled tau protein are the pathological hallmark CTE, and inhibiting aggregation or disassociating has been considered promising avenue to prevent treat CTE. Caffeine (CA) well-known psychostimulant can be found coffee, tea, soft drinks. In vitro experiments revealed that CA could effectively inhibit wild-type disassemble preformed fibrils. However, atomic effect underlying molecular mechanisms remain largely elusive. this study, we performed multitude replica exchange with solute tempering 2 (REST2) conventional dynamics (CMD) simulations 43.8 μs total on models without CA, including third fourth microtubule-binding repeats (R3-R4) monomer CTE-related R3-R4 protofibril fibril. results prominently β-sheet formation disrupt structure protofibril, inducing adopt loosely packed extended conformations. H-bonding π-π stacking interactions drove binding monomer, while hydrophobic made an extra contribution protofibril. Strikingly, stably bind cavity which might occupy space entering cofactor. What more, destabilized fibril played role reversing liquid-to-solid phase transition (LSPT) tau. Our study systematically uncovered atomic-level aggregation, offers theoretical foundation for design drugs

Язык: Английский

Процитировано

0