ceb52763f03c97b18267ea374edd0aa6 pmc_30042316.pdf 0d6495c5b188d4356a499d772b43708516f93954 pmc_30042316.pdf ca209a1ebb7133cd9d54c24acbe27f060aad581876590369fd5c656bb6d9172b pmc_30042316.pdf Title: Protein Environment: A Crucial Triggering Factor in Josephin Domain Aggregation: The Role of 2,2,2-Trifluoroethanol Subject: The protein ataxin-3 contains a polyglutamine stretch that triggers amyloid aggregation when it is expanded beyond a critical threshold. This results in the onset of the spinocerebellar ataxia type 3. The protein consists of the globular N-terminal Josephin domain and a disordered C-terminal tail where the polyglutamine stretch is located. Expanded ataxin-3 aggregates via a two-stage mechanism: first, Josephin domain self-association, then polyQ fibrillation. This highlights the intrinsic amyloidogenic potential of Josephin domain. Therefore, much effort has been put into investigating its aggregation mechanism(s). A key issue regards the conformational requirements for triggering amyloid aggregation, as it is believed that, generally, misfolding should precede aggregation. Here, we have assayed the effect of 2,2,2-trifluoroethanol, a co-solvent capable of stabilizing secondary structures, especially -helices. By combining biophysical methods and molecular dynamics, we demonstrated that both secondary and tertiary JD structures are virtually unchanged in the presence of up to 5% 2,2,2-trifluoroethanol. Despite the preservation of JD structure, 1% of 2,2,2-trifluoroethanol suffices to exacerbate the intrinsic aggregation propensity of this domain, by slightly decreasing its conformational stability. These results indicate that in the case of JD, conformational fluctuations might suffice to promote a transition towards an aggregated state without the need for extensive unfolding, and highlights the important role played by the environment on the aggregation of this globular domain. Keywords: ataxin-3; josephin domain; 2,2,2-trifluoroethanol; amyloid aggregation; molecular dynamics; protein-cosolvent interaction Author: Cristina Visentin, Susanna Navarro, Gianvito Grasso, Maria Elena Regonesi, Marco Agostino Deriu, Paolo Tortora and Salvador Ventura Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.15 CreationDate: Tue Jul 24 09:40:58 2018 CEST ModDate: Tue Jul 24 09:40:58 2018 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 17 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3512836 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- FBPZLW+URWPalladioL-Ital Type 1 Custom yes yes no 99 0 URJBAJ+URWPalladioL-Bold Type 1 Custom yes yes no 100 0 OXBKYU+TeXGyreAdventor-Bold Type 1 Custom yes yes no 101 0 LSWNHN+URWPalladioL-Roma Type 1 Custom yes yes no 102 0 TDYSNF+VnURWPalladioL 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Annotation: Subtype: Link Rect: 473, 262, 509, 273 Annotation: Subtype: Link Rect: 76, 220, 166, 251 Annotation: Subtype: Link Rect: 234, 215, 424, 226 Checksum: dad05ca2 Type: CRC32 Checksum: ceb52763f03c97b18267ea374edd0aa6 Type: MD5 Checksum: 0d6495c5b188d4356a499d772b43708516f93954 Type: SHA-1 Checksum: ca209a1ebb7133cd9d54c24acbe27f060aad581876590369fd5c656bb6d9172b Type: SHA-256