{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T17:06:19Z","timestamp":1780506379385,"version":"3.54.1"},"reference-count":87,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T00:00:00Z","timestamp":1769644800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T00:00:00Z","timestamp":1769644800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:p>\n                    Spinocerebellar ataxia type 3 is a debilitating neurodegenerative disorder driven by the pathological aggregation of ataxin\u20103 (Atx3), a deubiquitinating enzyme with a cysteine\u2010rich catalytic domain and an expandable polyglutamine (polyQ) tract. While the role of polyQ expansion in Atx3 aggregation is well documented, the influence of redox conditions on its self\u2010assembly remains underexplored. Here, we demonstrate that reducing agents and a redox environment critically modulate Atx3 aggregation by regulating disulfide bond formation within the Josephin domain. We demonstrate that dithiothreitol (DTT), through progressive oxidation, promotes the formation of non\u2010native and disulfide\u2010linked conformers, which may serve as nucleation centers for fibril formation. In contrast, tris(2\u2010carboxyethyl)phosphine preserves cysteine residues of Atx3 in the reduced state and inhibits aggregation, but concomitantly promotes cleavage of full\u2010length Atx3. Furthermore, we identify a previously underappreciated role for 4,5\u2010dihydroxy\u20101,2\u2010dithiane, the DTT oxidation product, in directly triggering Atx3 aggregation. We also demonstrate that running aggregation assays at 50\u2009\u00b0C circumvents the redox dependency of Atx3 aggregation, thereby streamlining the aggregation process and enabling the use of a simplified, robust platform for medium\u2010 to high\u2010throughput screening of aggregation modulators. These findings provide new insights into the redox\u2010dependent modulation of Atx3 aggregation and highlight critical considerations for\n                    <jats:italic>in vitro<\/jats:italic>\n                    aggregation assays of cysteine\u2010rich proteins, with broad implications for therapeutic strategies targeting cysteine\u2010rich, aggregation\u2010prone proteins in neurodegenerative diseases. Although our study focuses on\n                    <jats:italic>in vitro<\/jats:italic>\n                    investigation, it suggests that redox dysregulation in cells could promote pathogenic aggregation of Atx3, reinforcing the link between cellular redox balance and polyglutamine disease progression.\n                  <\/jats:p>","DOI":"10.1111\/febs.70426","type":"journal-article","created":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T14:21:56Z","timestamp":1769696516000},"page":"3376-3398","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Redox environment modulates aggregation of ataxin\u20103\n                    <i>in\u00a0vitro<\/i>\n                    \u2014 Implications for drug screening of cysteine\u2010rich proteins"],"prefix":"10.1111","volume":"293","author":[{"given":"Martyna","family":"Podlasiak","sequence":"first","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto  Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular, Universidade do Porto  Portugal"},{"name":"ICBAS \u2013 Instituto de Ci\u00eancias Biom\u00e9dicas Abel Salazar  Porto Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"Sollazzo","sequence":"additional","affiliation":[{"name":"Structural Biology and Biophysics Unit, Fondazione Ri.MED  Palermo Italy"},{"name":"Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (STEBICEF) University of Palermo  Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisa","family":"Monaca","sequence":"additional","affiliation":[{"name":"Structural Biology and Biophysics Unit, Fondazione Ri.MED  Palermo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raffaele","family":"Sabbatella","sequence":"additional","affiliation":[{"name":"Structural Biology and Biophysics Unit, Fondazione Ri.MED  Palermo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Agnese","family":"Morando","sequence":"additional","affiliation":[{"name":"Structural Biology and Biophysics Unit, Fondazione Ri.MED  Palermo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oleg","family":"Chertkov","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto  Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular, Universidade do Porto  Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Puglisi","sequence":"additional","affiliation":[{"name":"Structural Biology and Biophysics Unit, Fondazione Ri.MED  Palermo Italy"},{"name":"Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (STEBICEF) University of Palermo  Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"Mascellino","sequence":"additional","affiliation":[{"name":"Structural Biology and Biophysics Unit, Fondazione Ri.MED  Palermo Italy"},{"name":"Department of Biological, Chemical and Pharmaceutical Sciences and Technologies (STEBICEF) University of Palermo  Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Fricano","sequence":"additional","affiliation":[{"name":"Department of Physics and Chemistry Emilio Segr\u00e8 (DiFC) University of Palermo  Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandra","family":"Macedo\u2010Ribeiro","sequence":"additional","affiliation":[{"name":"i3S \u2013 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade, Universidade do Porto  Portugal"},{"name":"IBMC \u2013 Instituto de Biologia Molecular e Celular, Universidade do Porto  Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0374-5852","authenticated-orcid":false,"given":"Caterina","family":"Alfano","sequence":"additional","affiliation":[{"name":"Structural Biology and Biophysics Unit, Fondazione Ri.MED  Palermo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,1,29]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng1194-221"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/ana.410390411"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/ana.410410408"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pneurobio.2011.11.006"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41572-019-0074-3"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg1691"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.6b00936"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-444-63233-3.00011-7"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2018.02.037"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4614-5434-2_7"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2023.102607"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s004010100364"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(00)80943-5"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.19-23-10338.1999"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/8.4.673"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/ana.10101"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1440-1819.2003.01102.x"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00401-010-0717-7"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/bpa.12412"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(03)00748-8"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0013695"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2004.07.131"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0506344102"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0501732102"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.211305198"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M601470200"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2007.02.058"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2011.02.056"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2004.09.065"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2005.08.061"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M114.044610"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-006-6097-4"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0604547103"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2005.09.083"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb.1570"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells11121969"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2023.115258"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M405799200"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.659532"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201707345"},{"key":"e_1_2_10_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jasms.2010.01.016"},{"key":"e_1_2_10_43_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-4658.2006.05419.x"},{"key":"e_1_2_10_44_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21238916"},{"key":"e_1_2_10_45_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2006.10.093"},{"key":"e_1_2_10_46_1","doi-asserted-by":"publisher","DOI":"10.1097\/WCO.0b013e3283168d1d"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1134108"},{"key":"e_1_2_10_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-10574-w"},{"key":"e_1_2_10_49_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00892a002"},{"key":"e_1_2_10_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/0307-4412(83)90048-1"},{"key":"e_1_2_10_51_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1994.1290"},{"key":"e_1_2_10_52_1","doi-asserted-by":"publisher","DOI":"10.1006\/abio.1999.4203"},{"key":"e_1_2_10_53_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202000092"},{"key":"e_1_2_10_54_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0078060"},{"key":"e_1_2_10_55_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4949"},{"key":"e_1_2_10_56_1","first-page":"138","article-title":"Domain unfolding of monoclonal antibody fragments revealed by non\u2010reducing SDS\u2010PAGE","volume":"16","author":"Kirley TL","year":"2018","journal-title":"Biochem Biophys Rep"},{"key":"e_1_2_10_57_1","doi-asserted-by":"publisher","DOI":"10.1155\/2013\/318319"},{"key":"e_1_2_10_58_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tox.2021.153049"},{"key":"e_1_2_10_59_1","doi-asserted-by":"publisher","DOI":"10.3389\/fnmol.2020.582488"},{"key":"e_1_2_10_60_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijms.2012.08.032"},{"key":"e_1_2_10_61_1","doi-asserted-by":"publisher","DOI":"10.1038\/nsb961"},{"key":"e_1_2_10_62_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep15231"},{"key":"e_1_2_10_63_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0602048103"},{"key":"e_1_2_10_64_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0902505106"},{"key":"e_1_2_10_65_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.macromol.0c00861"},{"key":"e_1_2_10_66_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201307481"},{"key":"e_1_2_10_67_1","doi-asserted-by":"publisher","DOI":"10.1002\/chin.200320173"},{"key":"e_1_2_10_68_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2015.08.027"},{"key":"e_1_2_10_69_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mrgentox.2013.06.023"},{"key":"e_1_2_10_70_1","doi-asserted-by":"publisher","DOI":"10.3389\/fneur.2017.00485"},{"key":"e_1_2_10_71_1","doi-asserted-by":"publisher","DOI":"10.1155\/2020\/1270256"},{"key":"e_1_2_10_72_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neurobiolaging.2021.07.014"},{"key":"e_1_2_10_73_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms23115938"},{"key":"e_1_2_10_74_1","doi-asserted-by":"publisher","DOI":"10.3390\/antiox13030375"},{"key":"e_1_2_10_75_1","doi-asserted-by":"publisher","DOI":"10.1080\/1028415X.2024.2352195"},{"key":"e_1_2_10_76_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejphar.2024.177187"},{"key":"e_1_2_10_77_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10858-004-4343-3"},{"key":"e_1_2_10_78_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2018.05.029"},{"key":"e_1_2_10_79_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms160818741"},{"key":"e_1_2_10_80_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.C300049200"},{"key":"e_1_2_10_81_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2007.06.063"},{"key":"e_1_2_10_82_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12154-009-0027-5"},{"key":"e_1_2_10_83_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbapap.2010.04.001"},{"key":"e_1_2_10_84_1","doi-asserted-by":"publisher","DOI":"10.1021\/ed101169t"},{"key":"e_1_2_10_85_1","doi-asserted-by":"publisher","DOI":"10.2174\/157341105774573956"},{"key":"e_1_2_10_86_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.20449"},{"key":"e_1_2_10_87_1","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2024.1478241"},{"key":"e_1_2_10_88_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-024-55360-1"}],"container-title":["The FEBS Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/febs.70426","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/febs.70426","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/febs.70426","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T16:11:40Z","timestamp":1780503100000},"score":1,"resource":{"primary":{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/10.1111\/febs.70426"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,29]]},"references-count":87,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2026,6]]}},"alternative-id":["10.1111\/febs.70426"],"URL":"https:\/\/doi.org\/10.1111\/febs.70426","archive":["Portico"],"relation":{"has-review":[{"id-type":"doi","id":"10.1111\/FEBS.70426\/v2\/review1","asserted-by":"object"},{"id-type":"doi","id":"10.1111\/FEBS.70426\/v1\/decision1","asserted-by":"object"},{"id-type":"doi","id":"10.1111\/FEBS.70426\/v2\/response1","asserted-by":"object"},{"id-type":"doi","id":"10.1111\/FEBS.70426\/v1\/review1","asserted-by":"object"},{"id-type":"doi","id":"10.1111\/FEBS.70426\/v1\/review2","asserted-by":"object"},{"id-type":"doi","id":"10.1111\/FEBS.70426\/v2\/decision1","asserted-by":"object"}]},"ISSN":["1742-464X","1742-4658"],"issn-type":[{"value":"1742-464X","type":"print"},{"value":"1742-4658","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,29]]},"assertion":[{"value":"2025-07-30","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-01-19","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-01-29","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}