{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T18:01:04Z","timestamp":1790791264604,"version":"4.1.0"},"reference-count":68,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T00:00:00Z","timestamp":1738022400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2025,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Amyloid fibril formation of \u03b1\u2010synuclein (\u03b1SN) is a hallmark of synucleinopathies. Although the previous studies have provided numerous insights into the molecular basis of \u03b1SN amyloid formation, it remains unclear how \u03b1SN self\u2010assembles into amyloid fibrils in vivo. Here, we show that \u03b1SN amyloid formation is accelerated in the presence of two macromolecular crowders, polyethylene glycol (PEG) (MW: ~10,000) and dextran (DEX) (MW: ~500,000), with a maximum at approximately 7% (w\/v) PEG and 7% (w\/v) DEX. Under these conditions, the two crowders induce a two\u2010phase separation of upper PEG and lower DEX phases with a small number of liquid droplets of DEX and PEG in PEG and DEX phases, respectively. Fluorescence microscope images revealed that the interfaces of DEX droplets in the upper PEG phase are the major sites of amyloid formation. We consider that the depletion interactions working in micro phase\u2010segregated state with DEX and PEG systems causes \u03b1SN condensation at the interface between solute PEG and DEX droplets, resulting in accelerated amyloid formation. Ultrasonication further accelerated the amyloid formation in both DEX and PEG phases, confirming the droplet\u2010dependent amyloid formation. Similar PEG\/DEX\u2010dependent accelerated amyloid formation was observed for amyloid \u03b2 peptide. In contrast, amyloid formation of \u03b2\n                    <jats:sub>2<\/jats:sub>\n                    \u2010microglobulin or hen egg white lysozyme with a native fold was suppressed in the PEG\/DEX mixtures, suggesting that the depletion interactions work adversely depending on whether the protein is unfolded or folded.\n                  <\/jats:p>","DOI":"10.1002\/pro.5163","type":"journal-article","created":{"date-parts":[[2025,1,29]],"date-time":"2025-01-29T00:42:01Z","timestamp":1738111321000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Accelerated amyloid fibril formation at the interface of liquid\u2013liquid phase\u2010separated droplets by depletion interactions"],"prefix":"10.1002","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3408-1980","authenticated-orcid":false,"given":"Keiichi","family":"Yamaguchi","sequence":"first","affiliation":[{"name":"Graduate School of Engineering 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