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Elucidating the molecular determinants governing this process is essential for understanding the pathological mechanisms of diseases and developing therapeutic strategies that target early\u2010stage aggregation. While previous studies have identified residues critical for \u03b1\u2010synuclein amyloid formation, the key residues and molecular drivers of its phase separation remain largely unexplored. Herein, multiscale simulations and experimental approaches are employed to uncover the molecular determinants dictating \u03b1\u2010synuclein phase separation and the pre\u2010solidification of its condensates. Seven motifs are identified that exhibit high \u03b2\u2010sheet propensity in the monomeric state of \u03b1\u2010synuclein and progressively increase in \u03b2\u2010sheet content during condensation. Notably, two C\u2010terminal motifs engage in a percolated network of intermolecular interactions through transient hydrogen bonds, contributing to the phase boundary properties. Deletion of these motifs reduces the phase separation ability of \u03b1\u2010synuclein, underscoring their essential roles in this process. Together, the findings reveal crucial phase separation hotspots and shed light on the molecular mechanism underlying \u03b1\u2010synuclein phase separation, offering significant insights and novel potential therapeutic targets for Parkinson's disease.<\/jats:p>","DOI":"10.1002\/advs.202511545","type":"journal-article","created":{"date-parts":[[2025,9,8]],"date-time":"2025-09-08T09:55:59Z","timestamp":1757325359000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Dissecting the Molecular Determinants of \u03b1\u2010synuclein Phase Separation and Condensate Aging: The Pivotal Role of \u03b2\u2010Sheet\u2010Rich Motifs"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2976-5547","authenticated-orcid":false,"given":"Yiming","family":"Tang","sequence":"first","affiliation":[{"name":"Department of Physics State Key Laboratory of Surface Physics and Key Laboratory for Computational Physical Science (Ministry of Education) Fudan University  2005 Songhu Road, Yangpu District Shanghai 200433 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jitao","family":"Wen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences  15 Datun Road, Chaoyang District Beijing 100101 China"},{"name":"University of the Chinese Academy of Sciences  19A Yuquan Road, Shijingshan District Beijing 100049 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongyuan","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Physics State Key Laboratory of Surface Physics and Key Laboratory for Computational Physical Science (Ministry of Education) Fudan University  2005 Songhu Road, Yangpu District Shanghai 200433 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yifei","family":"Yao","sequence":"additional","affiliation":[{"name":"Department of Physics State Key Laboratory of Surface Physics and Key Laboratory for Computational Physical Science (Ministry of Education) Fudan University  2005 Songhu Road, Yangpu District Shanghai 200433 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaoshuai","family":"He","sequence":"additional","affiliation":[{"name":"Key Laboratory of Biomacromolecules CAS Center for Excellence in Biomacromolecules Institute of Biophysics Chinese Academy of Sciences  15 Datun Road, Chaoyang District Beijing 100101 China"},{"name":"University of the Chinese Academy of Sciences  19A Yuquan Road, Shijingshan District Beijing 100049 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiyuan","family":"Zeng","sequence":"additional","affiliation":[{"name":"Department of Physics State Key Laboratory of Surface Physics and Key Laboratory for Computational Physical Science (Ministry of Education) Fudan University  2005 Songhu Road, Yangpu District Shanghai 200433 China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuewei","family":"Dong","sequence":"additional","affiliation":[{"name":"Center for Soft Condensed Matter Physics and Interdisciplinary Research &amp; 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