Biophysical Chemistry, Journal Year: 2021, Volume and Issue: 281, P. 106736 - 106736
Published: Dec. 6, 2021
Language: Английский
Biochemistry, Journal Year: 2021, Volume and Issue: 60(48), P. 3676 - 3696
Published: Aug. 25, 2021
Liquid–liquid phase separation (LLPS) is a crucial phenomenon for the formation of functional membraneless organelles. However, LLPS also responsible protein aggregation in various neurodegenerative diseases such as amyotrophic lateral sclerosis, Alzheimer's disease, and Parkinson's disease (PD). Recently, several reports, including ours, have shown that α-synuclein (α-Syn) undergoes subsequent liquid-to-solid transition, which leads to amyloid fibril formation. how environmental (and experimental) parameters modulate α-Syn remains elusive. Here, we show vitro strongly dependent on presence salts, allows charge neutralization at both terminal segments therefore promotes hydrophobic interactions supportive LLPS. Using purification methods experimental conditions, showed, depending upon either spontaneous (instantaneous) or delayed Furthermore, delineate kinetics liquid droplet (i.e., critical concentration time) relative can be modulated by salt/counterion concentration, pH, surface, PD-associated multivalent cations, N-terminal acetylation, are all known regulate vitro. Together, our observations suggest transition could pathological, triggered only under disease-associated conditions (high and/or promoting self-assembly). This study will significantly improve understanding molecular mechanisms transition.
Language: Английский
Citations
114
Trends in Neurosciences, Journal Year: 2022, Volume and Issue: 46(2), P. 153 - 166
Published: Dec. 23, 2022
Language: Английский
Citations
97
Journal of Molecular Biology, Journal Year: 2022, Volume and Issue: 435(1), P. 167713 - 167713
Published: July 3, 2022
Language: Английский
Citations
83
Journal of Molecular Biology, Journal Year: 2023, Volume and Issue: 435(5), P. 167971 - 167971
Published: Jan. 21, 2023
In the past almost 15 years, we witnessed birth of a new scientific field focused on existence, formation, biological functions, and disease associations membraneless bodies in cells, now referred to as biomolecular condensates. Pioneering studies from several laboratories [reviewed in1, 2, 3] supported model wherein condensates associated with diverse processes form through process phase separation. These other findings that followed have revolutionized our understanding how biomolecules are organized space time within cells perform myriad including cell fate determination, signal transduction, endocytosis, regulation gene expression protein translation, RNA metabolism. Further, formed aberrant transitions been numerous human diseases, prominently neurodegeneration cancer. While some cases, rigorous evidence supports links between formation separation many others such less robustly supported, which has led rightful scrutiny generality roles biology disease.4, 5, 6, 7 During week-long workshop March 2022 at Telluride Science Research Center (TSRC) Telluride, Colorado, ∼25 scientists addressed key questions surrounding field. Herein, present insights gained these discussions, addressing topics including, systems, normal states, their applications synthetic biology, potential for therapeutically targeting
Language: Английский
Citations
53
Trends in Neurosciences, Journal Year: 2023, Volume and Issue: 46(4), P. 293 - 306
Published: Jan. 30, 2023
Neuronal communication crucially relies on exocytosis of neurotransmitters from synaptic vesicles (SVs) which are clustered at synapses. To ensure reliable neurotransmitter release, synapses need to maintain an adequate pool SVs all times. Decades research have established that by synapsin 1, abundant SV-associated phosphoprotein. The classical view postulates crosslinked in a scaffold protein-protein interactions between synapsins and their binding partners. Recent studies shown cluster via liquid-liquid phase separation (LLPS), thus providing new framework for the organization synapse. We discuss evidence SVs, emphasizing emerging questions related its regulation, specificity, reversibility.
Language: Английский
Citations
50
Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)
Published: Oct. 23, 2023
Neuronal transmission relies on the regulated secretion of neurotransmitters, which are packed in synaptic vesicles (SVs). Hundreds SVs accumulate at boutons. Despite being held together, highly mobile, so that they can be recruited to plasma membrane for their rapid release during neuronal activity. However, how such confinement corroborates with motility remains unclear. To bridge this gap, we employ ultrafast single-molecule tracking (SMT) reconstituted system native and living neurons. synapsin 1, most abundant protein, form condensates liquid-like properties. In these condensates, 1 movement is slowed both short (i.e., 60-nm) long several hundred-nm) ranges, suggesting SV-synapsin interaction raises overall packing condensate. Furthermore, two-color SMT super-resolution imaging axons demonstrate drives accumulation Even intrinsically-disordered fragment was sufficient restore SV pattern triple knock-out animals. Thus, condensation guarantee reliable SVs, allowing formation mesoscale domains synapses vivo.
Language: Английский
Citations
43
Biophysical Reports, Journal Year: 2021, Volume and Issue: 1(1), P. 100011 - 100011
Published: July 30, 2021
The material properties of biomolecular condensates have been suggested to play important biological and pathological roles. Despite the rapid increase in number biomolecules identified that undergo liquid-liquid phase separation, quantitative studies direct measurements resulting severely lagging behind. Here, we develop a micropipette-based technique uniquely, our knowledge, allows quantifications both surface tension viscosity condensates, independent labeling surface-wetting effects. We demonstrate accuracy versatility this by measuring LAF-1 RGG domains polymer-based aqueous two-phase system. further confirm using established condensate fusion fluorescence recovery after photobleaching assays. anticipate will be widely applicable resolve several limitations regarding current approaches.
Language: Английский
Citations
85
Journal of Neuroinflammation, Journal Year: 2022, Volume and Issue: 19(1)
Published: July 2, 2022
Deposition of amyloid beta (Aβ) and hyperphosphorylated tau along with glial cell-mediated neuroinflammation are prominent pathogenic hallmarks Alzheimer's disease (AD). In recent years, impairment autophagy has been identified as another important feature contributing to AD progression. Therefore, the potential activator spermidine, a small body-endogenous polyamine often used dietary supplement, was assessed on Aβ pathology neuroinflammation.Oral treatment prone AD-like APPPS1 mice spermidine reduced neurotoxic soluble decreased AD-associated neuroinflammation. Mechanistically, single nuclei sequencing revealed microglia be main target spermidine. This population characterized by increased AXL levels expression genes implicated in cell migration phagocytosis. A subsequent proteome analysis isolated confirmed anti-inflammatory cytoskeletal effects mice. primary astrocytes, spermidine-induced subsequently affected TLR3- TLR4-mediated inflammatory processes, phagocytosis motility. Interestingly, regulated neuroinflammatory response beyond transcriptional control interfering assembly inflammasome.Our data highlight that holds enhance degradation counteract glia-mediated pathology.
Language: Английский
Citations
61
Journal of Biological Chemistry, Journal Year: 2021, Volume and Issue: 298(1), P. 101469 - 101469
Published: Dec. 4, 2021
α-Synuclein (α-Syn) is the major protein component of Lewy bodies, a key pathological feature Parkinson's disease (PD). The manganese ion Mn2+ has been identified as an environmental risk factor PD. However, it remains unclear how regulates α-Syn aggregation. Here, we discovered that Mn2+accelerates amyloid aggregation through regulation phase separation. We found not only promotes liquid-to-solid transition but also directly induces soluble monomers to form solid-like condensates. Interestingly, lipid membrane integrated into condensates during Mn2+-induced transition; however, preformed Mn2+/α-syn can recruit lipids surface In addition, this largely facilitate Although do fuse, our results demonstrated they could existing Furthermore, observed chelator reverses stage. after maturation, becomes irreversible. These findings demonstrate facilitates accelerate formation aggregates and provide new insights for targeting separation in PD treatment. (α-Syn), cytoplasmic abundant presynaptic nerve terminals, implicated (PD), dementia with other synucleinopathies (1Baba M. Nakajo S. Tu P.H. Tomita T. Nakaya K. Lee V.M. Trojanowski J.Q. Iwatsubo Aggregation alpha-synuclein bodies sporadic bodies.Am. J. Pathol. 1998; 152: 879-884PubMed Google Scholar, 2Maroteaux L. Campanelli J.T. Scheller R.H. Synuclein: A neuron-specific localized nucleus terminal.J. Neurosci. 1988; 8: 2804-2815Crossref PubMed 3Bernal-Conde L.D. Ramos-Acevedo R. Reyes-Hernández M.A. Balbuena-Olvera A.J. Morales-Moreno I.D. Argüero-Sánchez Schüle B. Guerra-Crespo Alpha-synuclein physiology pathology: perspective on cellular structures organelles.Front. 2019; 13: 1399Crossref Scopus (41) Scholar). While monomer unstructured potential (4Fauvet Mbefo M.K. Fares M.B. Desobry C. Michael Ardah M.T. Tsika E. Coune P. Prudent Lion N. Eliezer D. Moore D.J. Schneider Aebischer El-Agnaf O.M. et al.α-Synuclein central nervous system from erythrocytes, mammalian cells, Escherichia coli exists predominantly disordered monomer.J. Biol. Chem. 2012; 287: 15345-15364Abstract Full Text PDF (387) 5Theillet F.X. Binolfi A. Bekei Martorana Rose H.M. Stuiver Verzini Lorenz van Rossum Goldfarb Selenko Structural disorder monomeric α-synuclein persists cells.Nature. 2016; 530: 45-50Crossref (525) 6Nguyen Derreumaux Structures intrinsically Aβ, tau proteins aqueous solution computer simulations.Biophys. 2020; 264: 106421Crossref (57) Scholar), multiple studies suggested forms α-helical tetramers physiologically resist (7Bartels Choi J.G. Selkoe occurs helically folded tetramer resists aggregation.Nature. 2011; 477: 107-110Crossref (847) 8Wang W. Perovic I. Chittuluru Kaganovich Nguyen L.T. Liao Auclair J.R. Johnson Landeru Simorellis A.K. Ju Cookson M.R. Asturias F.J. Agar J.N. Webb B.N. al.A construct dynamic tetramer.Proc. Natl. Acad. Sci. U. 108: 17797-17802Crossref (346) involved neurotransmitter release many critical trafficking processes. its physiological function completely understood (9Burré Sharma Tsetsenis Buchman V. Etherton Südhof T.C. SNARE-complex assembly vivo vitro.Science. 2010; 329: 1663-1667Crossref (1119) PD, oligomeric fibrillar intermolecular assemblies (10Nguyen Ramamoorthy Sahoo B.R. Zheng Faller Straub J.E. Dominguez Shea Dokholyan N.V. De Simone Ma Nussinov Najafi Ngo S.T. Loquet al.Amyloid oligomers: joint experimental/computational Alzheimer's disease, type II diabetes, amyotrophic lateral sclerosis.Chem. Rev. 2021; 121: 2545-2647Crossref (141) 11Agerschou E.D. Schützmann M.P. Reppert Wördehoff M.M. Shaykhalishahi H. Buell Hoyer β-Turn exchanges segment 44-TKEG-47 reveal high sequence fidelity requirements fibril elongation.Biophys. 269: 106519Crossref (5) neurites cytoplasm neurons are mainly composed aggregated (12Spillantini M.G. Schmidt M.L. Jakes Goedert alpha-Synuclein bodies.Nature. 1997; 388: 839-840Crossref (5862) 13Nemani Lu Berge Nakamura Onoa Chaudhry F.A. Nicoll R.A. Edwards Increased expression reduces by inhibiting synaptic vesicle reclustering endocytosis.Neuron. 65: 66-79Abstract (735) toxic, numerous disclose oligomers fibrils causes toxicity (14Karpinar D.P. Balija Kügler Opazo F. Rezaei-Ghaleh Wender Kim H.Y. Taschenberger G. Falkenburger B.H. Heise Kumar Riedel Fichtner Voigt Braus G.H. al.Pre-fibrillar variants impaired beta-structure increase neurotoxicity models.EMBO 2009; 28: 3256-3268Crossref (341) 15Volpicelli-Daley L.A. Luk K.C. Patel T.P. Tanik S.A. Riddle D.M. Stieber Meaney D.F. Exogenous induce body pathology leading dysfunction neuron death.Neuron. 72: 57-71Abstract (899) Many pathway alternations, such oxidative stress, mitochondrial dysfunction, endoplasmic reticulum autophagy-lysosomal have linked cytotoxic effects (15Volpicelli-Daley 16Fields C.R. Bengoa-Vergniory Wade-Martins Targeting therapy disease.Front. Mol. 12: 299Crossref (96) 17Wong Y.C. Krainc neurodegeneration: Mechanism therapeutic strategies.Nat. Med. 2017; 23: 1-13Crossref (395) 18Guiney S.J. Adlard P.A. Lei Mawal C.H. Bush A.I. Finkelstein D.I. Ayton Fibrillar depends functional lysosomes.J. 295: 17497-17513Abstract (13) aggregations proteins, tau, IAPP, FUS, TDP-43, studied vitro decades 19Nguyen Sterpone Pouplana Campanera J.M. Dimerization mechanism Alzheimer Aβ(40) peptides: content intrapeptide-stabilized conformations A2V A2T heterozygous dimers retards formation.J. Phys. 120: 12111-12126Crossref (34) 20Ivanova M.I. Lin Y. Y.H. Biophysical processes underlying cross-seeding implications pathology.Biophys. 106507Crossref Protein targets amyloid-related 21Doig Inhibition small molecules.Curr. Opin. Struct. 2015; 30: 50-56Crossref (212) 22Savitt Jankovic disease: Progress towards development disease-modifying therapeutics.Drugs. 79: 797-810Crossref (45) addition α-Syn, contain components, including various organelles (20Ivanova 23Shahmoradian S.H. Lewis Genoud Hench Moors T.E. Navarro P.P. Castaño-Díez Schweighauser Graff-Meyer Goldie K.N. Sütterlin Huisman Ingrassia Gier Rozemuller A.J.M. al.Lewy consists crowded membranes.Nat. 22: 1099-1109Crossref (331) factors, lipids, coexist affect accumulation For example, activity glucocerebrosidase, lysosomal glycoprotein affects levels (24Muñoz S.S. Petersen Marlet F.R. Kücükköse Galvagnion interplay between human models disease.Biophys. 273: 106534Crossref (16) Recent Raman spectral imaging revealed colocalized cells (25Watson M.D. Flynn J.D. J.C. (13)C(2)H(15)N-labeled cells.Biophys. 106528Crossref (8) while regulations previously (26Galvagnion Meisl Michaels Vendruscolo Knowles Dobson C.M. Lipid vesicles trigger stimulating primary nucleation.Nat. 11: 229-234Crossref (358) 27Giehm Svergun Otzen D.E. Vestergaard Low-resolution structure disrupting oligomer accumulates fibrillation.Proc. 3246-3251Crossref (194) 28Kurochka A.S. Yushchenko D.A. Bouř Shvadchak V.V. Influence membranes aggregation.ACS 825-830Crossref (10) Metal ions Cu2+, Zn2+, concentrated brain bind (29González Arcos-López König Quintanar Menacho Márquez Outeiro T.F. Fernández C.O. Effects post-translational modifications metal binding.J. Neurochem. 150: 507-521Crossref (37) C-terminal domain which contains Asp Glu residues, interact di- trivalent 30Binolfi Rasia R.M. Bertoncini C.W. Ceolin Zweckstetter Griesinger Jovin T.M. Interaction divalent reveals differences: link structure, binding specificity fibrillation enhancement.J. Am. Soc. 2006; 128: 9893-9901Crossref (262) detailed information each metal-α-Syn still due lack high-resolution data. Manganese essential (31Pfalzer A.C. Bowman A.B. Relationships biology neurological disease.Curr. Environ. Health Rep. 4: 223-228Crossref 32Tinkov A.A. Paoliello M.M.B. Mazilina A.N. Skalny A.V. Martins Voskresenskaya O.N. Aaseth Santamaria Notova S.V. Tsatsakis Aschner Molecular manganese-induced neurotoxicity: five-year update.Int. 4646Crossref rise utility industry led increased pollution. imbalance homeostasis cause neurodegenerative diseases (33Guilarte T.R. New perspectives behavioral, neuroimaging, neuropathological humans non-human primates.Front. Aging 2013; 5: 23Crossref (136) 34Miah Ijomone Okoh C.O.A. O.K. Akingbade G.T. Ke Krum da Cunha Jr., Akinyemi Aranoff Antunes Soares overexposure health.Neurochem. Int. 135: 104688Crossref (35) Specially, epidemiological investigations indicate excessive exposure increases (34Miah 35Gorell C.C. Rybicki B.A. Peterson E.L. Kortsha G.X. Brown G.G. Richardson R.J. Occupational manganese, copper, lead, iron, mercury zinc disease.Neurotoxicology. 1999; 20: 239-247PubMed 36Lucchini R.G. Martin C.J. Doney B.C. From manganism parkinsonism: conceptual model based evolution exposure.Neuromolecular 311-321Crossref (125) Multiple occupational welding, mining, related industrial activities associated Parkinsonism (37Racette McGee-Minnich Moerlein S.M. Mink J.W. Videen T.O. Perlmutter J.S. Welding-related Clinical features, treatment, pathophysiology.Neurology. 2001; 56: 8-13Crossref (306) 38Jiang Y.M. Mo X.A. Du F.Q. Fu X. Zhu X.Y. Gao Xie J.L. F.L. Pira Effective treatment parkinsonism p-aminosalicylic acid: case 17-year follow-up study.J. Occup. 48: 644-649Crossref (108) 39Racette Tabbal S.D. Jennings Good Evanoff Prevalence relationship large sample Alabama welders.Neurology. 2005; 64: 230-235Crossref (150) 40Rutchik Jiang How does neurologist assess welders steelworkers movement disorder? An international team's experiences Guanxi, China part II.J. 54: 1562-1564Crossref (7) 41Rutchik China, I.J. 1432-1434Crossref (9) 42Racette Criswell S.R. Lundin J.I. Hobson Seixas Kotzbauer P.T. Zhang Sheppard Checkoway welding exposure.Neurotoxicology. 33: 1356-1361Crossref (105) 43Mortimer J.A. Borenstein A.R. Nelson L.M. Associations Parkinson Review meta-analysis.Neurology. 1174-1180Crossref (63) 44Al-Lozi Nielsen Hershey Birke Racette Cognitive control workers exposed manganese-containing fume.Am. Ind. 60: 181-188Crossref (15) Long-time level air may deficits cognitive (45Bowler Kornblith E.S. Gocheva Colledge Bollweg Beseler C.L. Wright Adams S.W. Lobdell D.T. Environmental air: functions.Neurotoxicology. 49: 139-148Crossref 46Carvalho C.F. Oulhote Martorelli Carvalho Menezes-Filho Argollo Abreu associations memory, executive functions, hyperactivity Brazilian children.Neurotoxicology. 2018; 69: 253-259Crossref (24) recent meta-analysis previous 22 significantly circulating patients, further demonstrating (47Du Liu M.Y. Pan Y.Z. Zhong Wei M.J. Association meta-analysis.Neurosci. Lett. 665: 92-98Crossref One induced (48Sun He Yang Liang Shi Ou injury via ATP-sensitive K(+) channels GABA receptors.Toxicol. 332: 164-170Crossref 49Liu Z.Q. Z.F. Cong Z. Li Deng Xu Manganese-induced overexpression aggravates damage repressing PINK1/parkin-mediated mitophagy.Food Toxicol. 112213Crossref (3) 50Verina Guilarte frontal cortex primates.Toxicol. 217: 177-183Crossref 51Cai Yao Chen Cao Shen Luo PC12 ERK activation.Brain Res. 1359: 201-207Crossref (47) 52Harischandra D.S. Rokad Neal Ghaisas Manne Sarkar Panicker Zenitsky Jin Huang Anantharam Kanthasamy A.G. prion-like cell-to-cell exosomal transmission α-synuclein.Sci. Signal. 12eaau4543Crossref (74) most these were cell culture, their relevance unknown. Biochemical showed shortens time (53Han J.Y. T.S. H.I. role Ca(2+) hard cations accelerated interfibrillar 1895Crossref (33) early stage elusive. protein–protein or protein–RNA condensed liquid droplets weak multivalent interactions, called liquid–liquid (LLPS). Proteins undergo LLPS, Tau protein, usually repetitive domains regions (IDR) (54Li Banjade Cheng H.C. Guo Llaguno Hollingsworth J.V. King Banani S.F. Russo P.S. Q.X. Nixon B.T. Rosen Phase transitions signalling proteins.Nature. 483: 336-340Crossref (1140) 55Conicella A.E. Dignon G.L. Zerze H.B. D'Ordine A.M. Rohatgi Ayala Mittal Fawzi N.L. TDP-43 tunes liquid-liquid function.Proc. 117: 5883-5894Crossref (102) 56Levone Lenzken S.C. Antonaci Maiser Rapp Conte Reber Mechtersheimer Ronchi Mühlemann O. Leonhardt Cardoso M.C. Ruepp Barabino S.M.L. FUS-dependent important DNA repair initiation.J. Cell 220e202008030Crossref 57Ambadipudi Biernat Mandelkow Liquid-liquid microtubule-binding repeats Alzheimer-related tau.Nat. Commun. 275Crossref (308) Numerous variety aggregation-prone initiate LLPS (58Wegmann Eftekharzadeh Tepper Zoltowska K.M. Bennett R.E. Dujardin Laskowski P.R. MacKenzie Kamath Commins Vanderburg Roe A.D. Fan Molliex Hernandez-Vega al.Tau aggregation.EMBO 37e98049Crossref (368) 59Tange Ishibashi Nakagaki Taguchi Kamatari Y.O. Ozawa Nishida full-length prion initiates conformational conversion vitro.J. 296: 100367Abstract (22) 60Pytowski Foley Vaux Jean diabetes-associated IAPP hydrogelation aggregation.Proc. 12050-12061Crossref 61Xing Nandakumar Kakinen Sun Davis P.C. Ding Amyloid under lens separation.J. 368-378Crossref (12) three IDRs low-complexity domains. reported undergoes aggregation, indicating be intermediate before (62Ray Singh Pandey Datta Mahato Panigrahi Navalkar Mehra Gadhe Chatterjee Sawner Maiti Bhatia nucleates separation.Nat. 705-716Crossref (147) 63Hardenberg Sinnige Casford Dada Poudel Robinson E.A. Fuxreiter Kaminksi Kaminski-Schierle G.S. Nollen E.A.A. Observation droplet state maturation body-like assemblies.J. 282-294PubMed 64Hoffmann Sansevrino Morabito Logan Vabulas Ulusoy Ganzella Milovanovic Synapsin alpha-synuclein.J. 433: 166961Crossref (20) work, examined whether LLPS. requirement crowding agents. efficiently transition, facilitating absence agents, bypassing step presence membrane. Lipids recruited Our proceed recruitment monomers. difficult disassemble mature aggregates, chelators reverse dimension understanding pathogenesis suggest target therapy. To study regulatory expressed (Fig. S1) conditions drives vitro. first labeled preparing EGFP-α-Syn fusion protein. unlabeled mixed at molar ratio 1:9 Using fluorescence microscopy, underwent relatively concentration S2) agent PEG-10000 S3). Ficoll 400, another macromolecular agent, promote although efficiency lower S4). By contrast, failed drive small-molecule ethylene glycol, supporting acts α-syn S5). Next, tested plays PEG, frequently used mimic environment, 200 μM formed typical smooth droplets. When included 2 mM Mn2+, morphology turned irregular 1A). dose-dependence experiments when was 400 above S6). Then performed recovery photobleaching (FRAP) examine intensity returned prebleaching 120 s laser bleaching, rapid fluidity inside 1, B C). contras
Language: Английский
Citations
60
Biochemical and Biophysical Research Communications, Journal Year: 2022, Volume and Issue: 603, P. 13 - 20
Published: Feb. 25, 2022
Language: Английский
Citations
40