{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T15:25:55Z","timestamp":1789226755851,"version":"build-2803163510"},"reference-count":64,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2025,6,16]],"date-time":"2025-06-16T00:00:00Z","timestamp":1750032000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy are disorders characterized by the presence of cytosolic \u03b1\u2010synuclein (SNCA) amyloids. The gene <jats:italic>SNCA<\/jats:italic> is alternatively spliced, generating three variants of SNCA, missing exon 3 (SNCA\u03943) or 5 (SNCA\u03945), or both exons (SNCA\u03943\u03945). Despite purported upregulation in disease states, their pathological relevance is ill\u2010defined. Here, we investigated the amyloid formation of alternatively spliced variants under physiological conditions. Aggregation kinetics, secondary structure, and fibril morphology of N\u2010terminally acetylated SNCA\u03943, SNCA\u03945, and SNCA\u03943\u03945 were assessed by thioflavin\u2010T fluorescence, circular dichroism spectroscopy, and transmission electron microscopy, respectively. Compared to SNCA, both SNCA\u03945 and SNCA\u03943\u03945 aggregate faster and adopt a more twisted fibril morphology, whereas SNCA\u03943 is more sensitive to solution conditions, exhibiting similar or modestly faster aggregation kinetics compared to SNCA. Cross\u2010seeding experiments using spliced\u2010variant fibrils and soluble SNCA showed that despite fibril morphological differences, SNCA\u03945 were competent seeds for SNCA, which is explained by their similar protease\u2010K resistant regions. Contrastingly, neither SNCA\u03943 nor SNCA\u03943\u03945 fibrils cross\u2010seed SNCA, indicating exon 3 (residues 41\u201354) is essential in modulating fibril structure. Notably, SNCA aggregation is stimulated by sub\u2010stoichiometric amounts of soluble SNCA\u03945 and SNCA\u03943\u03945, but not SNCA\u03943, suggesting that exon 5 (residues 103\u2013130) is more important in modulating aggregation kinetics. Taken together, we propose that alternatively spliced variants are pathogenic by exacerbating aggregation of the main SNCA isoform.<\/jats:p>","DOI":"10.1002\/pro.70195","type":"journal-article","created":{"date-parts":[[2025,6,16]],"date-time":"2025-06-16T13:56:23Z","timestamp":1750082183000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Amyloid formation of alternatively spliced variants of \u03b1\u2010synuclein"],"prefix":"10.1002","volume":"34","author":[{"given":"Daniel Q.","family":"SanGiovanni","sequence":"first","affiliation":[{"name":"Laboratory of Protein Conformation and Dynamics, Biochemistry and Biophysics Center National Heart, Lung, and Blood Institute, National Institutes of Health  Bethesda Maryland USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan P.","family":"McGlinchey","sequence":"additional","affiliation":[{"name":"Laboratory of Protein Conformation and Dynamics, 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