The role of RNA in the nanoscale organization of alpha-synuclein phase separation DOI Creative Commons
Sabrina Zappone, Eleonora Perego, Jakob Rupert

et al.

Deleted Journal, Journal Year: 2025, Volume and Issue: 2(2)

Published: March 29, 2025

Abstract The cellular accumulation of α-synuclein (aS) aggregates is a hallmark several neurodegenerative diseases. Recent studies suggest that the aberrant transition monomeric aS into solid-like may occur through an intermediate liquid-like state, where protein partitions between dense and dilute phases. Although not typically recognized as RNA-binding protein, it can bind RNA under aggregation conditions, but its impact on phases remains unexplored. Employing combination fluorescence spectroscopy techniques, we investigated mobility in both presence RNA. Our analysis revealed formation nanoclusters involved initiating phase separation uncovered heterogeneity within phase, discovering molecules exist two distinct states. Additionally, demonstrated induces morphological changes promotes liquid-to-solid phase. These findings underscore active role modulating transitions.

Language: Английский

Mass photometric detection and quantification of nanoscale α-synuclein phase separation DOI
Soumik Ray, Thomas O. Mason, Lars Boyens‐Thiele

et al.

Nature Chemistry, Journal Year: 2023, Volume and Issue: 15(9), P. 1306 - 1316

Published: June 19, 2023

Language: Английский

Citations

68
HDX–MS finds that partial unfolding with sequential domain activation controls condensation of a cellular stress marker DOI Creative Commons
Ruofan Chen, Hendrik Glauninger, Darren N. Kahan

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2024, Volume and Issue: 121(13)

Published: March 21, 2024

Eukaryotic cells form condensates to sense and adapt their environment [S. F. Banani, H. O. Lee, A. Hyman, M. K. Rosen, Nat. Rev. Mol. Cell Biol. 18 , 285–298 (2017), Yoo, C. Triandafillou, D. Drummond, J. Chem. 294 7151–7159 (2019)]. Poly(A)-binding protein (Pab1), a canonical stress granule marker, condenses upon heat shock or starvation, promoting adaptation [J. Riback et al. 168 1028–1040.e19 (2017)]. The molecular basis of condensation has remained elusive due dearth techniques probe structure directly in condensates. We apply hydrogen–deuterium exchange/mass spectrometry investigate the mechanism Pab1’s condensation. four RNA recognition motifs (RRMs) undergo different levels partial unfolding condensation, changes are similar for thermal pH stresses. Although structural heterogeneity is observed, ability MS describe populations allows us identify which regions contribute condensate’s interaction network. Our data yield picture stress-triggered we term sequential activation ( Fig. 1 A ), wherein each RRM becomes activated at temperature where it partially unfolds associates with other likewise RRMs condensate. Subsequent association dictated more by underlying free energy surface than specific interactions, an effect refer as thermodynamic specificity. study represents advance elucidating interactions that drive Furthermore, our findings demonstrate how can use specificity perform acute response multiple stresses, potentially general stress-responsive proteins.

Language: Английский

Citations

20
Direct Observation of “Elongated” Conformational States in α‐Synuclein upon Liquid‐Liquid Phase Separation DOI Creative Commons

Daniele Ubbiali,

Marta Fratini, Lolita Piersimoni

et al.

Angewandte Chemie International Edition, Journal Year: 2022, Volume and Issue: 61(46)

Published: Sept. 17, 2022

Abstract α‐Synuclein (α‐syn) is an intrinsically disordered protein (IDP) that undergoes liquid‐liquid phase separation (LLPS), fibrillation, and forms insoluble intracellular Lewy bodies in neurons, which are the hallmark of Parkinson's Disease (PD). Neurotoxicity precedes formation aggregates might be related to α‐syn LLPS. The molecular mechanisms underlying early stages LLPS still elusive. To obtain structural insights into upon LLPS, we take advantage cross‐linking/mass spectrometry (XL–MS) introduce innovative approach, termed COMPASS (COMPetitive PAiring StatisticS). In this work, show conformational ensemble shifts from a “hairpin‐like” structure towards more “elongated” states We critical initial establish novel mass spectrometry‐based approach will aid solve open questions biology.

Language: Английский

Citations

43
Long non‐coding RNAs as key regulators of neurodegenerative protein aggregation DOI Creative Commons

Qi Xu,

Dan Liu,

Ling‐Qiang Zhu

et al.

Alzheimer s & Dementia, Journal Year: 2025, Volume and Issue: 21(2)

Published: Feb. 1, 2025

Abstract The characteristic events in neurodegenerative diseases (NDDs) encompass protein misfolding, aggregation, accumulation, and their related cellular dysfunction, synaptic function loss. While distinct proteins are implicated the pathological processes of different NDDs, process misfolding aggregation remains notably similar across various conditions. Specifically, undergo into beta‐folded (β‐folded) conformation, resulting formation insoluble amyloid proteins. Despite advancements comprehending certain facets this intricate remain incompletely elucidated. In recent years, concept that long non‐coding RNAs (lncRNAs) contribute to has gained recognition. LncRNAs influence aggregates by facilitating overexpression through regulation gene transcription translation, inhibiting degradation via lysosomal autophagic pathways, targeting aberrant modifications phase transitions A better understanding relationship between lncRNAs is an important step dissecting underlying molecular mechanisms will discovery new therapeutic targets strategies. Highlights NDDs marked leading dysfunction loss function. being involved β‐folded conformations forming consistent role attention, as they regulate inhibit degradation, target modifications. Understanding link crucial for uncovering developing targets.

Language: Английский

Citations

1
神经退行性疾病相关淀粉样蛋白的液-液相分离 DOI

Yun-Yi Luo,

Yanmei Li

Chinese Science Bulletin (Chinese Version), Journal Year: 2025, Volume and Issue: 70(7), P. 850 - 859

Published: Feb. 12, 2025

Citations

1
Experimental Evidence for Phosphorylation-Driven Allosteric Regulation of Alpha Synuclein Function DOI Creative Commons

Ashlyn N. Dollar,

Ian Webb

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 26, 2025

Phosphorylation of serine 129 (pS129) in the intrinsically disordered protein alpha synuclein has long been associated with neurodegenerative disease. In past several years, functional relevance pS219 uncovered by electrophysiology, immunoprecipitation, and proteomics as intricately connected neurotransmitter release synaptic vesicle (SV) cycling. Unexpectedly, binding to SNARE complex proteins VAMP-2 synapsin only occurs phosphorylation-competent synuclein. The domain shown be residues 96-110, which does not include phosphorylated residue, hinting at allosteric regulation protein-protein interactions pS129. Within this study, cross-linking, covalent labeling, collision induced unfolding pS129 - well an additional encountered form brain, oxidized-M1, M5, M116, M127 are studied utilizing tandem mass spectrometry. Collision gives a fingerprint structures' relative compactness stabilities various conformations. Covalent labeling identifies solvent accessible reveals hydrophobicity (or hydrophilicity) their microenvironment, while cross-linking maps proximity residue pairs. combination unfolding, show unequivocally that phosphorylated-S129 results more stable, compact form. Our provide evidence extensively folded amphipathic region interacts strongly domain. phosphorylation-induced folding likely tunes other SVs membranes.

Language: Английский

Citations

1
Distinct Effects of Familial Parkinson’s Disease-Associated Mutations on α-Synuclein Phase Separation and Amyloid Aggregation DOI Creative Commons

Bingkuan Xu,

Fengshuo Fan,

Yunpeng Liu

et al.

Biomolecules, Journal Year: 2023, Volume and Issue: 13(5), P. 726 - 726

Published: April 23, 2023

The Lewy bodies and neurites are key pathological hallmarks of Parkinson’s disease (PD). Single-point mutations associated with familial PD cause α-synuclein (α-Syn) aggregation, leading to the formation neurites. Recent studies suggest α-Syn nucleates through liquid–liquid phase separation (LLPS) form amyloid aggregates in a condensate pathway. How PD-associated affect LLPS its correlation aggregation remains incompletely understood. Here, we examined effects five identified PD, A30P, E46K, H50Q, A53T, A53E, on α-Syn. All other mutants behave similarly wild-type (WT) α-Syn, except that E46K mutation substantially promotes condensates. mutant droplets fuse WT recruit monomers into their droplets. Our showed A53T accelerated In contrast, A53E retarded during liquid-to-solid transition. Finally, observed formed condensates cells, whereas apparently promoted These findings reveal have divergent phase-separated condensates, providing new insights pathogenesis mutations.

Language: Английский

Citations

21
Illuminating Intrinsically Disordered Proteins with Integrative Structural Biology DOI Creative Commons

Rachel Evans,

Sravani Ramisetty,

Prakash Kulkarni

et al.

Biomolecules, Journal Year: 2023, Volume and Issue: 13(1), P. 124 - 124

Published: Jan. 7, 2023

Intense study of intrinsically disordered proteins (IDPs) did not begin in earnest until the late 1990s when a few groups, working independently, convinced community that these ‘weird’ could have important functions. Over past two decades, it has become clear IDPs play critical roles multitude biological phenomena with prominent examples including coordination signaling hubs, enabling gene regulation, and regulating ion channels, just to name few. One contributing factor delayed appreciation IDP functional significance is experimental difficulty characterizing their dynamic conformations. The combined application multiple methods, termed integrative structural biology, emerged as an essential approach understanding phenomena. Here, we review some recent applications biology philosophy IDPs.

Language: Английский

Citations

19
Challenges in studying the liquid-to-solid phase transitions of proteins using computer simulations DOI Creative Commons
Beata Szała-Mendyk, Tien M. Phan, Priyesh Mohanty

et al.

Current Opinion in Chemical Biology, Journal Year: 2023, Volume and Issue: 75, P. 102333 - 102333

Published: May 31, 2023

Language: Английский

Citations

17
Sequence-Dependent Conformational Transitions of Disordered Proteins During Condensation DOI Creative Commons
Jiahui Wang, Dinesh Sundaravadivelu Devarajan,

Young C. Kim

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 12, 2024

Intrinsically disordered proteins (IDPs) can form biomolecular condensates through phase separation. It is recognized that the conformation of IDPs in dense and dilute phases as well at interfaces critically impact resulting properties associated with their functionality. However, a comprehensive understanding conformational transitions during condensation remains elusive. In this study, we employ coarse-grained polyampholyte model, comprising an equal number oppositely charged residues-glutamic acid lysine-whereby conformations behavior be readily tuned by altering protein sequence. By manipulating sequence patterns from perfectly alternating to block-like, obtain chains ideal-like semi-compact structures phase, while chain approximately ideal chain, irrespective performing simulations different concentrations, find assemble small oligomeric clusters accompanied gradual swelling individual chains. We further demonstrate these findings are applicable several naturally occurring involved formation biological condensates. Concurrently, delve deeper into within condensate, revealing interface show strong dependence, but remain more collapsed than those bulk-like phase. This study addresses critical gaps our knowledge IDP function

Language: Английский

Citations

8