Heterotypic electrostatic interactions control complex phase separation of tau and prion into multiphasic condensates and co-aggregates

Proceedings of the National Academy of Sciences, Journal Year: 2023, Volume and Issue: 120(2)

Published: Jan. 3, 2023

Biomolecular condensates formed via phase separation of proteins and nucleic acids are thought to perform a wide range critical cellular functions by maintaining spatiotemporal regulation organizing intracellular biochemistry. However, aberrant transitions implicated in multitude human diseases. Here, we demonstrate that two neuronal proteins, namely tau prion, undergo complex coacervation driven domain-specific electrostatic interactions yield highly dynamic, mesoscopic liquid-like droplets. The acidic N-terminal segment interacts electrostatically with the polybasic intrinsically disordered prion protein (PrP). We employed unique combination time-resolved tools encompass several orders magnitude timescales ranging from nanoseconds seconds. These studies unveil an intriguing symphony molecular events associated formation heterotypic comprising ephemeral, domain-specific, short-range nanoclusters. Our results reveal these can be tuned RNA stoichiometry-dependent manner resulting reversible, multiphasic, immiscible, ternary different morphologies core-shell nested This system exhibits typical three-regime behavior reminiscent other membraneless organelles including nucleolar condensates. also show upon aging, tau:PrP droplets gradually convert into solid-like co-assemblies sequestration persistent intermolecular interactions. vibrational Raman conjunction atomic force microscopy multi-color fluorescence imaging presence amorphous amyloid-like co-aggregates maturation. findings provide mechanistic underpinnings overlapping neuropathology involving PrP highlight broader biological role physiology disease.

Language: Английский

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Non-classical crystallization in soft and organic materials

Nature Reviews Materials, Journal Year: 2024, Volume and Issue: 9(4), P. 229 - 248

Published: Feb. 6, 2024

Language: Английский

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Protein misfolding and amyloid nucleation through liquid–liquid phase separation

Chemical Society Reviews, Journal Year: 2024, Volume and Issue: 53(10), P. 4976 - 5013

Published: Jan. 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.

Language: Английский

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The role of biomolecular condensates in protein aggregation

Nature Reviews Chemistry, Journal Year: 2024, Volume and Issue: 8(9), P. 686 - 700

Published: Aug. 12, 2024

Language: Английский

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Principles Governing the Phase Separation of Multidomain Proteins

Biochemistry, Journal Year: 2022, Volume and Issue: 61(22), P. 2443 - 2455

Published: July 8, 2022

A variety of membraneless organelles, often termed "biological condensates", play an important role in the regulation cellular processes such as gene transcription, translation, and protein quality control. On basis experimental theoretical investigations, liquid–liquid phase separation (LLPS) has been proposed a possible mechanism for origin biological condensates. LLPS requires multivalent macromolecules that template formation long-range, intermolecular interaction networks results condensates with defined composition material properties. Multivalent interactions driving exhibit wide range modes from highly stereospecific to nonspecific involve both folded disordered regions. Multidomain proteins serve suitable promoting achieving disparate functions due their potential regulation. Here, we aim highlight influence domain architecture interdomain on multidomain First, general principles underlying these are illustrated examples predominantly associated nucleic acid binding control contain Next, showcase how properties regions can be leveraged engineer constructs form desired assembly functional Finally, need improvements coarse-grained computational models provide molecular-level insights into conjunction efforts.

Language: Английский

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Recent Advances in Microgels: From Biomolecules to Functionality

Small, Journal Year: 2022, Volume and Issue: 18(34)

Published: July 5, 2022

Abstract The emerging applications of hydrogel materials at different length scales, in areas ranging from sustainability to health, have driven the progress design and manufacturing microgels. Microgels can provide miniaturized, monodisperse, regulatable compartments, which be spatially separated or interconnected. These microscopic novel opportunities for generating biomimetic cell culture environments are thus key advances modern biomedical research. evolution physical chemical properties has, furthermore, highlighted potentials microgels context science bioengineering. This review describes recent research fabrication, characterization, generated biomolecular building blocks. A enabling technology allowing tailoring is their synthesis through microfluidic technologies, this paper highlights these impact on expanding physicochemical parameter space accessible using finally discusses roles that play liquid–liquid phase separation, micromechanics, biosensors, regenerative medicine.

Language: Английский

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The Role of Post-Translational Modifications on the Structure and Function of Tau Protein

Journal of Molecular Neuroscience, Journal Year: 2022, Volume and Issue: 72(8), P. 1557 - 1571

Published: March 24, 2022

Language: Английский

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Protein structural transitions critically transform the network connectivity and viscoelasticity of RNA-binding protein condensates but RNA can prevent it

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: Sept. 29, 2022

Abstract Biomolecular condensates, some of which are liquid-like during health, can age over time becoming gel-like pathological systems. One potential source loss properties ageing RNA-binding protein condensates is the progressive formation inter-protein β -sheets. To bridge microscopic understanding between accumulation -sheets and modulation FUS hnRNPA1 condensate viscoelasticity, we develop a multiscale simulation approach. Our method integrates atomistic simulations with sequence-dependent coarse-grained modelling that exhibit time. We reveal notably increase viscosity but does not transform phase diagrams. Strikingly, network molecular connections within drastically altered, culminating in gelation when strong fully percolates. However, high concentrations RNA decelerate emergence study uncovers kinetic factors explaining how trigger liquid-to-solid transitions suggests mechanism to slow such down.

Language: Английский

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Liquid–liquid phase separation of amyloid-β oligomers modulates amyloid fibrils formation

Journal of Biological Chemistry, Journal Year: 2023, Volume and Issue: 299(3), P. 102926 - 102926

Published: Jan. 20, 2023

Soluble amyloid-β oligomers (AβOs) are proposed to instigate and mediate the pathology of Alzheimer's disease, but mechanisms involved not clear. In this study, we reported that AβOs can undergo liquid–liquid phase separation (LLPS) form liquid-like droplets in vitro. We determined exhibited an α-helix conformation a membrane-mimicking environment SDS. Importantly, SDS is capable reconfiguring assembly different induce their LLPS. Moreover, found droplet formation was promoted by strong hydrated anions weak cations, suggesting hydrophobic interactions play key role mediating AβOs. Finally, observed LLPS further promote Aβ amyloid fibrils, which be modulated (−)-epigallocatechin gallate. Our study highlights as important entity protein liquid-to-solid transition reveals regulatory underlying aggregation, may relevant pathological process disease. The aberrant aggregation proteins from soluble state insoluble fibrils linked pathogenesis neurodegenerative diseases, including disease (AD), amyotrophic lateral sclerosis, Parkinson's type 2 diabetes (1Chiti F. Dobson C.M. Protein misfolding, functional amyloid, human disease.Annu. Rev. Biochem. 2006; 75: 333-366Crossref PubMed Scopus (5243) Google Scholar, 2Chiti formation, disease: summary progress over last decade.Annu. 2017; 86: 27-68Crossref (1536) 3Iadanza M.G. Jackson M.P. Hewitt E.W. Ranson N.A. Radford S.E. A new era for understanding structures disease.Nat. Mol. Cell Biol. 2018; 19: 755-773Crossref (487) Scholar). addition misfolding many these disease-associated dynamic condensates, fundamental normal cellular processing (4Wegmann S. Eftekharzadeh B. Tepper K. Zoltowska K.M. Bennett R.E. Dujardin et al.Tau initiate tau aggregation.EMBO J. 37: e98049Crossref (509) 5Kanaan N.M. Hamel C. Grabinski T. Combs Liquid-liquid induces pathogenic conformations vitro.Nat. Commun. 2020; 11: 2809Crossref (130) 6Ray Singh N. Kumar R. Patel Pandey Datta D. al.α-Synuclein nucleates through separation.Nat. Chem. 12: 705-716Crossref (268) 7Pytowski L. Lee C.F. Foley A.C. Vaux D.J. Jean Liquid–liquid II diabetes-associated IAPP initiates hydrogelation aggregation.Proc. Natl. Acad. Sci. U. A. 117: 12050-12061Crossref (38) For example, Alzheimer's-associated forms liquid at physiological levels vitro has function increase local concentration tubulin nucleate microtubule bundles (8Hernández-Vega Braun M. Scharrel Jahnel Wegmann Hyman B.T. al.Local nucleation into condensed phase.Cell Rep. 20: 2304-2312Abstract Full Text PDF (199) Similarly, also been during α-synuclein yields hydrogels associated with toxicity (6Ray Islet polypeptide undergoes catalyzed hydrophobic–hydrophilic interfaces (e.g., air-water interface vitro) LLPS-driven (7Pytowski addition, fused sarcoma (FUS), heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), TAR DNA-binding 43 kDa (TDP-43) inclusions sclerosis patients participating stress granule via regulate metabolism RNA (9Molliex Temirov Coughlin Kanagaraj A.P. Kim H.J. al.Phase low complexity domains promotes drives fibrillization.Cell. 2015; 163: 123-133Abstract (1518) 10Zbinden Pérez-Berlanga De Rossi P. Polymenidou Phase diseases: disturbance force.Dev. Cell. 55: 45-68Abstract (142) 11Patel H.O. Jawerth Maharana Hein M.Y. al.A ALS FUS accelerated mutation.Cell. 162: 1066-1077Abstract (1575) reversible micro-sized droplets. intrinsically disordered regions, domains, repetition motifs prone facilitate separation. driven multivalent primarily include electrostatic (12Brangwynne C.P. Tompa Pappu R.V. Polymer physics intracellular transitions.Nat. Phys. 899-904Crossref (822) 13Dignon G.L. Best R.B. Mittal Biomolecular separation: Molecular Driving forces macroscopic properties.Annu. 71: 53-75Crossref (200) Scholar), π–π cation–π (14Vernon R.M. Chong P.A. Tsang T.H. Bah Farber al.Pi-Pi contacts overlooked feature separation.eLife. 7e31486Crossref (388) (15Murthy Dignon Kan Y. Zerze G.H. Parekh S.H. al.Molecular liquid−liquid low-complexity domain.Nat. Struct. 2019; 26: 637-648Crossref (294) Scholar) etc. actively contributes maturation deposition, maturate less-dynamic finally yield irreversible (11Patel 16Gui X. Luo Li Zhou H. Qin Z. Liu al.Structural basis amyloids hnRNPA1 elucidates assembly.Nat. 10: 2006Crossref (115) 17Wen Hong Krainer G. Yao Q.-Q. Knowles T.P.J. Wu al.Conformational expansion condensates aggregation.J. Am. Soc. 2021; 143: 13056-13064Crossref (44) mature formed vitro, amyloid-like filaments share similar morphology structure assemblies diseases 18Burke K.A. Janke A.M. Rhine C.L. Fawzi N.L. Residue-by-Residue view granules bind C-terminal domain polymerase II.Mol. 60: 231-241Abstract (532) 19Lin Protter D.S.W. Rosen M.K. Parker Formation phase-separated RNA-binding proteins.Mol. 208-219Abstract (980) indicating early event neurodegeneration (20Alberti Dormann Genet. 53: 171-194Crossref (360) Therefore, targeting block while maintaining functions will lead potential therapeutics diseases. Amyloid initially self-assemble various associate pathway (3Iadanza This normally regarded sequential oligomerization discrete. Assemblies intermediates exert cytotoxicity (21Lee S.J.C. Nam E. Savelieff Lim M.H. Towards oligomers: characterization, mechanisms, inhibitors.Chem. 46: 310-323Crossref Oligomerization related toxic like (5Kanaan N-terminal TDP-43 demonstrated enhance full-length (22Wang Conicella A.E. Schmidt H.B. Martin Rhoads S.N. Reeb A.N. single phosphomimic disrupts polymerization, separation, splicing.EMBO e97452Crossref (210) However, whether exhibit behavior how it links poorly understood. work, high-throughput (HiPPS) profiling method (23Li Gu exploring parameter space liquid-liquid separation.Cell 2022; 3100764PubMed were prepared reformed enabled Meanwhile, redirected fibrils. finding provides insights mechanism regulates transition. Low molecular weight (MW) globular Aβ42 (Aβ42Os) 0.2% (w/v) previously described (24Barghorn Nimmrich V. Striebinger Krantz Keller Janson al.Globular beta-peptide1-42 oligomer - homogenous stable neuropathological disease.J. Neurochem. 2005; 95: 834-847Crossref (488) During Aβ42Os preparation, dimethyl sulfoxide (DMSO) solvent introduced dissolve peptide film (for detailed information, see Experimental procedures). To exclude interference DMSO on stability examine effects dissolution buffer conditions species, dissolved either (Protocol I) or DMSO-free condition NaOH solution II). electrophoresis, transmission electron microscopy (TEM) imaging technique, CD measurement performed evaluate properties MW, morphology, secondary structure. results (Fig. S1, A–D) two preparation protocols species. Spherical diameter ∼10 nm TEM 1A). band MW ∼18 displayed Tricine-SDS-PAGE, corresponding tetramer 1B). After large number screenings HiPPS 50 mM Tris/HCl pH 8.5 aqueous containing 1 M ammonium sulfate. occurred within wide range concentrations between 5 μM ∼ 200 μM, where (Figs. 1C S1E). depends salt 1F). Both synthetic recombinant monomeric peptides applied suggest sources under above S1F). Aβ42Os, labeled FITC-Aβ42 synthesized prepare fluorescent observation. FITC-Aβ42Os mixed unlabeled molar ratio 1:25. small spontaneously one large-size seconds fluorescence microscope 1D), indicates dense phase. Additionally, mobility assessed recovery after photobleaching (FRAP) measurement. signal recovered up 60% bleaching area (∼2 μm circle diameter) inside 1E), suggests rapid rearrangement molecules relatively FRAP reflects property due high viscosity nature phenomenon exclusively featured mentioned another methods employed, off-pathway Aβ-derived diffusible aggregates 25Dahlgren K.N. Manelli Stine W.B. Baker L.K. Krafft G.A. LaDu M.J. Oligomeric fibrillar species differentially affect neuronal viability.J. 2002; 277: 32046-32053Abstract (1256) on-pathway generated low-salt, low-temperature (26Ahmed Davis Aucoin Sato Ahuja Aimoto conversion neurotoxic amyloid-β1–42 fibrils.Nat. 2010; 17: 561-567Crossref (880) induced cannot Intriguingly, upon SDS, rapidly types S2). 0.1% 2% size increased increased, propose faster fusion 2A). investigate possible reason essential carried out native-PAGE measurements analyze structural changes produced disc-shaped pentamers adopt loosely aggregated strands accessible turns conformation. Upon break down trimers tetramers our composed presence shown spectra 2B). Besides, result showed higher 66 2C). Altogether, indicate tertiary speculate reconfigures assembly, required remodulation chains residues Aβ42. NMR recorded obtain information folding dilute dispersed 1D 1H spectrum (black Fig. 2D) unfolded, random coil peptides. Poor chemical shift dispersion intensity (red attributed coexistence two-state unfolding transitions. there exchange unfolded monomer well-folded oligomers. fraction total signal, whereas causes line broadening leading poor spectral resolution. Due both phase, decrease (blue observed, reflecting occurrence coprecipitating Besides sought find factors influence Aβ42Os. independent temperature temperatures S3A). (range 3.5 8.5) had negligible its S3B). Through systematical screenings, salts strongly (such citrate3-) weakly cations NH4+). contrast, NO3-) Mg2+) 3A). explained Hofmeister phenomena (27Cacace Landau E.M. Ramsden J.J. hofmeister series: interfacial phenomena.Q. Biophys. 1997; 30: 241-277Crossref (879) Series shows denaturation effectiveness ions sequences interacting water molecules, thus enables disables hydration shell. shell decreases free interact proteins, vital stability. More specifically, NH4+ salting-out ability rather than protein–solvent interactions. comparison relative later positions Mg2+ NO3-), solubility weakening strength 5% 1,6-hexanediol, suppression 3B). It validates These findings coincidence previous studies other general rule govern (28Krainer Welsh T.J. Joseph J.A. Espinosa J.R. Wittmann de Csilléry al.Reentrant condensate stabilized non-ionic interactions.Nat. 1085Crossref As modulate regulating interactions, explored metal (Mg2+, Zn2+, Al3+) At beginning LLPS, Al3+ stabilize state, tiny detected Zn2+. noteworthy mention show slower fluidity might change rigidity probably coincubation quiescently 37 °C 24 h, bulk cases, retarding S4) directly oligomeric state. involving serve driving force (29Sawaya M.R. Hughes Rodriguez Riek Eisenberg D.S. expanding family: structure, stability, function, pathogenesis.Cell. 184: 4857-4873Abstract (83) correlates fibrillization, incubated days. controls, monitored alone. SDS-trapped almost no 4 days S5A). sulfate chloride, h 4A thioflavin-T (ThT) assay used monitor validate ThT gradual enhancement incubation, adopting cross-β No significant nonphase-separated 4B). light microscope. By alone did any 4C). imply redirect formation. confirmed dot blot anti-amyloid OC antibody S5B). chemiluminescence remarkably alone, amount gallate (EGCG) inhibitor influences droplet's applying equivalent EGCG only became smaller gradually shrank 4D). 1:50 (peptide:EGCG), confocal Based research, could alter prevent (30Ehrnhoefer D.E. Bieschke Boeddrich Herbst Masino Lurz al.EGCG redirects amyloidogenic polypeptides unstructured, oligomers.Nat. 2008; 15: 558-566Crossref (1166) 31Ahmed VanSchouwen Jafari Ni Ortega Melacini (−)-Epigallocatechin gallate-induced nontoxic remodeling oligomers.J. 139: 13720-13734Crossref (72) suppose alters disfavoring Misfolded oligomerization, critical biochemical processes development biomolecular 32Marzahn Marada Nourse Kenrick Zhao al.Higher-order localization SPOP speckles.EMBO 2016; 35: 1254-1275Crossref (131) 33Babinchak W.M. Haider Dumm B.K. Sarkar Surewicz Choi J.-K. al.The domain.J. 294: 6306-6317Abstract (162) Recent RNA-mediated protein, presents progression model high-order (34Ash P.E.A. Lei Shattuck Boudeau Carlomagno Medalla al.TIA1 potentiates generation tau.Proc. 118e2014188118Crossref (49) typically most responsible neurotoxicity assembled cells even absence predominant hallmarks AD, do always correlate well neuron disability 35Glabe C.G. Structural classification 283: 29639-29643Abstract (662) 36Benilova I. Karran Strooper emperor need clothes.Nat. Neurosci. 2012; 349-357Crossref (1525) Thus, investigations relationship pathobiology generate valency lower energy barrier demix components distinct phases. Yet, exact still unknown. Here, AD-related underwent Sequence-based analysis (37Chu Sun Q. Xu Zhang Lai al.Prediction separating using machine learning.BMC Bioinform. 23: 72Crossref probability prediction outcome consistent experimental

Language: Английский

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Interaction Dynamics of Intrinsically Disordered Proteins from Single-Molecule Spectroscopy

Annual Review of Biophysics, Journal Year: 2023, Volume and Issue: 52(1), P. 433 - 462

Published: Feb. 8, 2023

Many proteins contain large structurally disordered regions or are entirely under physiological conditions. The functions of these intrinsically (IDPs) often involve interactions with other biomolecules. An important emerging effort has thus been to identify the molecular mechanisms IDP and how they differ from textbook notions biomolecular binding for folded proteins. In this review, we summarize versatile tool kit single-molecule fluorescence spectroscopy can aid investigation conformationally heterogeneous highly dynamic systems. We discuss experimental observables that be employed enable complexes probed on timescales nanoseconds hours. Key insights include diverse structural properties bound IDPs kinetic facilitated by disorder, such as fly-casting; disorder-mediated encounter complexes; competitive substitution via ternary complexes, which enables rapid dissociation even high-affinity complexes. also links aggregation, liquid-liquid phase separation, cellular processes, well current technical advances further expand scope spectroscopy.

Language: Английский

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