Protein misfolding and amyloid nucleation through liquid–liquid phase separation

Chemical Society Reviews, Год журнала: 2024, Номер 53(10), С. 4976 - 5013

Опубликована: Янв. 1, 2024

Protein misfolding and amyloid aggregation, linked to neurodegenerative diseases, can result from liquid–liquid phase separation (LLPS) a subsequent liquid-to-solid transition. This represents LLPS as generic mechanism in nucleation.

Язык: Английский

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From a Droplet to a Fibril and from a Fibril to a Droplet: Intertwined Transition Pathways in Highly Dynamic Enzyme-Modulated Peptide-Adenosine Triphosphate Systems

Journal of the American Chemical Society, Год журнала: 2024, Номер 146(9), С. 6045 - 6052

Опубликована: Фев. 23, 2024

Many cellular coassemblies of proteins and polynucleotides facilitate liquid–liquid phase separation (LLPS) the subsequent self-assembly disease-associated amyloid fibrils within liquid droplets. Here, we explore dynamics coupled conformational transitions model adenosine triphosphate (ATP)-binding peptides, ACC1–13Kn, consisting potent amyloidogenic fragment insulin's A-chain (ACC1–13) merged with oligolysine segments various lengths (Kn, n = 16, 24, 40). The ATP-stabilized is preceded by LLPS for peptides sufficiently long segments. two-component droplets are in dynamic equilibria free ATP monomeric which makes them susceptible to ATP-hydrolyzing apyrase ACC1–13Kn-digesting proteinase K. Both enzymes capable rapid disassembly fibrils, producing either monomers peptide (apyrase) or released together cleaved-off (proteinase K). In latter case, enzyme-sequestered Kn form co-released ATP, resulting an unusual fibril-to-droplet transition. support highly nature aggregate-monomer equilibria, addition superstoichiometric amounts ACC1–13Kn-ATP coaggregate causes its disassembly. Our results show that droplet state not merely intermediate on pathway aggregate but may also constitute final a complex protein misfolding scenario rich degraded fragments incompetent transition again into fibrils.

Язык: Английский

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Oncogenic p53 triggers amyloid aggregation of p63 and p73 liquid droplets

Communications Chemistry, Год журнала: 2024, Номер 7(1)

Опубликована: Сен. 16, 2024

P53 Phase separation is crucial towards amyloid aggregation and p63 p73 have enhanced expression in tumors. This study examines the phase behaviors of p53, p63, p73. Here we show that unlike DNA-binding domain p53 (p53C), p63C p73C undergo separation, but do not form amyloids under physiological temperatures. Wild-type mutant p53C droplets at 4°C aggregates 37 °C with properties. Mutant promotes amyloid-like states p73C, recruiting them into membraneless organelles. Amyloid conversion supported by thioflavin T Congo red binding, increased light scattering, circular dichroism. Full-length (or p73C) co-transfection shows reduced fluorescence recovery after photobleaching. Heparin inhibits prion-like induced p53C. These findings highlight role initiating p73, opening avenues for targeting cancer therapy.

Язык: Английский

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Amyloid Aggregation and Liquid–Liquid Phase Separation from the Perspective of Phase Transitions

The Journal of Physical Chemistry B, Год журнала: 2023, Номер 127(28), С. 6241 - 6250

Опубликована: Июль 6, 2023

Amyloid aggregation describes the aberrant self-assembly of peptides into ordered fibrils characterized by cross-β spine cores and is associated with many neurodegenerative diseases Type 2 diabetes. Oligomers, populated during early stage aggregation, are found to be more cytotoxic than mature fibrils. Recently, amyloidogenic have been reported undergo liquid–liquid phase separation (LLPS)─a biological process important for compartmentalization biomolecules in living cells─prior fibril formation. Understanding relationship between LLPS amyloid especially formation oligomers, essential uncovering disease mechanisms mitigating toxicity. In this Perspective, available theories models first briefly reviewed. By drawing analogies gas, liquid, solid phases thermodynamics, a diagram protein monomer, droplet, states separated coexistence lines can inferred. Due high free energy barrier fibrillization kinetically delaying seeds out droplets, "hidden" monomer-droplet line extends phase. then described as equilibration from initial "out-of-equilibrium" state homogeneous solution monomers final equilibrium stable coexisting and/or droplets via metastable or intermediates. The oligomers also discussed. We suggest that droplet should considered future studies which may help better understand develop therapeutic strategies mitigate

Язык: Английский

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The emerging role of ATP as a cosolute for biomolecular processes

Biological Chemistry, Год журнала: 2023, Номер 404(10), С. 897 - 908

Опубликована: Сен. 1, 2023

ATP is an important small molecule that appears at outstandingly high concentration within the cellular medium. Apart from its use as a source of energy and metabolite, there increasing evidence for functions cosolute biomolecular processes. Owned to solubilizing kosmotropic triphosphate hydrophobic adenine moieties, versatile can interact with biomolecules in various ways. We here three models categorize these interactions apply them review recent studies. focus on impact solubility, folding stability phase transitions. This leads us possible implications therapeutic interventions neurodegenerative diseases.

Язык: Английский

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Liquid–Liquid Phase Separation (LLPS)-Driven Fibrilization of Amyloid-β Protein

ACS Chemical Neuroscience, Год журнала: 2023, Номер 14(19), С. 3655 - 3664

Опубликована: Сен. 18, 2023

Amyloid-β [Aβ(1-40)] aggregation into a fibrillar network is one of the major hallmarks Alzheimer's disease (AD). Recently, few studies reported that polyphosphate (polyP), an anionic biopolymer participates in various cellular physiological processes humans, induces fibrilization many amyloidogenic proteins [ 2020 Disease Facts and Figures; John Wiley Sons Inc., 2020; Tanzi, R. E.; Bertram, L. Cell 2005, 120, 545-555; Selkoe, D. J. Proc. Natl. Acad. Sci. U.S.A. 1995, 275, 630-631; Rambaran, N.; Serpell, C. Prion 2008, 2, 112-117]. However, role polyP Aβ(1-40) underlying mechanism are unclear. In this study, we report experimental investigations on kinetics Aβ(1-40). It found exhibits dual effect depending upon pH value. At = 7 (neutral), inhibits amyloid dose-dependent manner similar to negatively charged nanoparticles. On contrary, at 3 (acidic), accelerates via liquid-liquid phase separation (LLPS), wherein protein-rich droplets contain mature fibrils. parameter space spanned by concentrations polyP, diagram constructed demark domain where LLPS observed 3. Characterization protein aggregates, secondary structure content cell viability presence aggregates discussed both values. This study reveals biopolymers can modulate kinetics, linked neurodegenerative diseases, their local pH.

Язык: Английский

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Coalescence-Driven Local Crowding Promotes Liquid-to-Solid-Like Phase Transition in a Homogeneous and Heterogeneous Droplet Assembly: Regulatory Role of Ligands

Langmuir, Год журнала: 2025, Номер unknown

Опубликована: Апрель 14, 2025

Liquid-to-solid-like phase transition (LSPT) of disordered proteins via metastable liquid-like droplets is a well-documented phenomenon in biology and linked to many pathological conditions including neurodegenerative diseases. However, very less known about the early microscopic events transient intermediates involved irreversible protein aggregation functional globular proteins. Herein, using range spectroscopic techniques, we show that LSPT protein, human serum albumin (HSA), exclusively driven by spontaneous coalescence involving various temporal manner. We interdroplet communication essential for both initial growth amorphous aggregates within individual droplets, which subsequently transform amyloid-like fibrils. Immobilized neither any nucleation nor upon aging. Moreover, found exchange materials with dilute dispersed has negligible influence on HSA. Our findings reveal interfacial properties effectively modulate feasibility kinetics HSA ligand binding, suggesting possible regulatory mechanism cells utilize control dynamics LSPT. Furthermore, dynamic heterogeneous droplet assembly two proteins, transferrin (Tf), an intriguing fused where solid-like phases coexist same droplet, eventually mixed fibrillar assembly. These insights not only highlight importance interactions behind biomolecules but also showcase its adverse effect structure function other crowded

Язык: Английский

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An aberrant fused in sarcoma liquid droplet of amyotrophic lateral sclerosis pathological variant, R495X, accelerates liquid–solid phase transition

Scientific Reports, Год журнала: 2024, Номер 14(1)

Опубликована: Апрель 17, 2024

Abstract Intracellular aggregation of fused in sarcoma (FUS) is associated with the pathogenesis familial amyotrophic lateral sclerosis (ALS). Under stress, FUS forms liquid droplets via liquid–liquid phase separation (LLPS). Two types wild-type LLPS exist equilibrium: low-pressure (LP-LLPS) and high-pressure (HP-LLPS); former dominates below 2 kbar latter over kbar. Although several disease-type variants have been identified, molecular mechanism underlying accelerated cytoplasmic granule formation ALS patients remains poorly understood. Herein, we report reversible two states irreversible liquid–solid transition, namely droplet aging, patient-type variant R495X using fluorescence microscopy ultraviolet–visible absorption spectroscopy combined perturbations pressure temperature. Liquid-to-solid transition was HP-LLPS than variant; arginine slowed aging at atmospheric conditions by inhibiting more selectively compared to that LP-LLPS. Our findings provide new insight into which readily aggregates. Targeting aberrantly formed (the state) proteins minimal impact on physiological functions could be a novel therapeutic strategy for LLPS-mediated protein diseases.

Язык: Английский

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Elucidating ATP’s Role as Solubilizer of Biomolecular Aggregate

eLife, Год журнала: 2024, Номер 13

Опубликована: Июль 10, 2024

Proteins occurring in significantly high concentrations cellular environments (over 100 mg/ml) and functioning crowded cytoplasm, often face the prodigious challenges of aggregation which are pathological hallmark aging critically responsible for a wide spectrum rising human diseases. Here, we combine joint-venture complementary wet-lab experiment molecular simulation to discern potential ability adenosine triphosphate (ATP) as solubilizer protein aggregates. We show that ATP prevents both condensation aggregation-prone intrinsically disordered Aβ40 promotes dissolution preformed Computer links ATP’s solubilizing role its modulate protein’s structural plasticity by unwinding conformation. is positioned superior biological aggregates over traditional chemical hydrotropes, potentially holding promises therapeutic interventions protein-aggregation-related Going beyond conventional activity energy currency, amphiphilic nature enables protein-specific interaction would enhance efficiency processes.

Язык: Английский

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Elucidating ATP’s Role as Solubilizer of Biomolecular Aggregate

Опубликована: Сен. 24, 2024

Proteins occurring in significantly high concentrations cellular environments (over 100 mg/mL) and functioning crowded cytoplasm, often face the prodigious challenges of aggregation which are pathological hallmark aging critically responsible for a wide spectrum rising human diseases. Here we combine joint-venture complementary wet-lab experiment molecular simulation to discern potential ability adenosine triphosphate (ATP) as solubilizer protein aggregates. We show that ATP prevents both condensation aggregation-prone intrinsically disordered Aβ40 promotes dissolution pre-formed Computer links ATP’s solubilizing role its modulate protein’s structural plasticity by unwinding conformation. is positioned superior biological aggregates over traditional chemical hydrotropes, potentially holding promises therapeutic interventions protein-aggregation related Going beyond conventional activity energy currency, amphiphilic nature enables protein-specific interaction would enhance efficiency processes.

Язык: Английский

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