Assessing amyloid fibrils and amorphous aggregates: A review

International Journal of Biological Macromolecules, Год журнала: 2025, Номер unknown, С. 143725 - 143725

Опубликована: Май 1, 2025

Protein misfolding and aggregation play a central role in the progression of neurodegenerative diseases such as Alzheimer's Parkinson's. These aggregates manifest either structured amyloid fibrils enriched β-sheet conformations or irregular amorphous with diverse morphologies. Understanding their formation, structure, behavior is critical for deciphering disease mechanisms developing targeted diagnostics therapeutics. This review presents an integrated overview both conventional advanced techniques used to detect, distinguish, structurally characterize these protein aggregates. It covers range spectroscopic spectrometric tools, fluorescence, Raman, mass spectrometry that facilitate aggregate identification. Microscopy methods, including atomic force electron microscopy, are highlighted morphological analysis. The also discusses situ detection strategies using fluorescent dyes, conformation-specific antibodies, enzymatic reporters, real-time imaging. Separation methods like centrifugation, electrophoresis, chromatography outlined alongside structural analysis tools X-ray diffraction. Furthermore, growing utility computational approaches artificial intelligence predicting propensities integrating biological data emphasized. By critically evaluating each method's capabilities limitations, this provides practical forward-looking resource researchers studying complex landscape aggregation.

Язык: Английский

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Thioflavin T: induced circular dichroism and density functional theory as promising approach in amyloid fibril assays

Applied Spectroscopy Reviews, Год журнала: 2025, Номер unknown, С. 1 - 39

Опубликована: Май 11, 2025

Язык: Английский

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Overexpression in Plasmodium falciparum of an intrinsically disordered protein segment of PfUT impairs the parasite’s proteostasis and reduces its growth rate

Frontiers in Cellular and Infection Microbiology, Год журнала: 2025, Номер 15

Опубликована: Май 13, 2025

The proteome of Plasmodium falciparum exhibits a marked propensity for aggregation. This characteristic results from the parasite’s AT-rich genome, which encodes numerous proteins with long asparagine-rich stretches and low structural complexity, lead to abundant intrinsically disordered regions. While this poses challenges parasite, protein aggregation may also serve functional roles, such as stress adaptation, could therefore be exploited by targeting it potential vulnerable spot in pathogen. Here, we overexpressed an aggregation-prone segment P. ubiquitin transferase ( Pf UTf), E3 ligase that has been previously demonstrated regulate stability parasite involved invasion, development drug metabolism. Overexpression UTf had evident phenotypic effects observed transmission electron microscopy confocal fluorescence microscopy, increased endogenous aggregation, disrupted proteostasis, caused significant growth impairment parasite. Combined dihydroartemisinin treatment, overexpression synergistic effect further compromised parasite´s viability, linking proteasome dysfunction. Changes distribution proteins, shown altered subcellular fluorescent pattern new investigational aggregated dye antiplasmodial compound YAT2150 overexpressing line, highlighted critical balance between responses, suggesting proteostasis-targeting therapies good antimalarial strategy.

Язык: Английский

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Thioflavin T Lasing Probe for Mucin Detection in Simulated Tears as a Targeting Strategy for Brain Tumors

ACS Chemical Neuroscience, Год журнала: 2025, Номер unknown

Опубликована: Май 15, 2025

A novel optical approach for a noninvasive detection of mucins in tear fluid is proposed, aiming at the early diagnosis brain tumors such as glioblastoma. Utilizing Thioflavin T (ThT) fluorescent probe, our study demonstrates that ThT selectively binds to (modeled by MUC3) DEMI water, artificial tears, and simulated tears. Steady-state time-resolved fluorescence spectroscopy reveal mucin binding induces significant enhancement prolonged emission lifetime, indicative restricted intramolecular rotation. Importantly, application Fabry-Pérot cavity lasing enabled resolution distinct spectral signatures ThT-mucin complex, including emergence dual peaks an increased threshold mucin-rich samples compared controls. These fingerprints provide compelling evidence specific interactions are not discernible with conventional techniques. Our findings highlight potential probe method sensitive, platform detecting offering promising strategy

Язык: Английский

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Cavity Lasing of Thioflavin T in the Condensed Phase for Discrimination between Surface Interaction and β-Sheet Groove Binding in Alzheimer-Linked Peptides

The Journal of Physical Chemistry Letters, Год журнала: 2024, Номер unknown, С. 9543 - 9547

Опубликована: Сен. 12, 2024

This study investigates the lasing effects in a Fabry-Perot cavity to discern binding interactions of thioflavin T (ThT) with various peptides associated Alzheimer's disease, including Aβ(1-42), KLVFFA, and diphenylalanine (FF) condensed phase. Utilizing kinetic measurements, research explores ThT emission enhancements due specific groove β-sheet structures highlights additional contributions from weak surface solvent-solute interactions. Lasing spectroscopy reveals lack transition FF system its native state an amyloid-like structure, challenging traditional assay interpretations. These findings show potential elucidating molecular basis amyloid fibril formation development diagnostic tools for amyloidogenic diseases.

Язык: Английский

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Quantification of Rapid Cooling of Glycerol/Water Solutions Based on Photoluminescence from Thioflavin T

The Journal of Physical Chemistry B, Год журнала: 2024, Номер 128(49), С. 12310 - 12324

Опубликована: Дек. 3, 2024

Rapid cooling to a solid state allows intermediates in chemical and biomolecular processes that occur solution near room temperature be trapped for subsequent measurements by magnetic resonance spectroscopies, electron microscopy, or other techniques. In time-resolved nuclear rapid freeze-quench paramagnetic studies, solutions are typically frozen spraying into cold bath onto metal surface. Although simulations suggest freezing on millisecond submillisecond time scales, direct experimental of rates have been elusive. Here, we describe method quantification based temperature-dependent photoluminescence from thioflavin T (ThT). our experiments, jet ThT glycerol/water, with 10.8 m/s velocity 30 μm diameter, freezes cold, rotating copper Images the surface indicate rate increases linearly over 0.45–6.2 range. At velocities greater than 3.8 m/s, cool 300 260 K 230 is less 100 μs 700 μs. The results do not agree quantitatively calculations which layer glycerol/water cools thermal conduction when suddenly brought contact Discrepancies between simplistic illustrate importance rates.

Язык: Английский

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Hemoglobin-derived amyloid fibrils: Fibrillization mechanisms and potential applications

Food Chemistry, Год журнала: 2024, Номер 470, С. 142671 - 142671

Опубликована: Дек. 29, 2024

Язык: Английский

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