1. Liquid–Liquid Phase Separation Enhances TDP-43 LCD Aggregation but Delays Seeded Aggregation
- Author
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Ludo Van Den Bosch, Frederic Rousseau, Emiel Michiels, Mathias De Decker, Peter Tompa, Sylvie Derclaye, Anna Bratek-Skicki, Philip Van Damme, David Alsteens, Joris Van Lindt, Donya Pakravan, Joost Schymkowitz, UCL - SST/LIBST - Louvain Institute of Biomolecular Science and Technology, Department of Bio-engineering Sciences, Faculty of Sciences and Bioengineering Sciences, and Structural Biology Brussels
- Subjects
Biochemistry & Molecular Biology ,Amyloid ,amyotrophic lateral sclerosis ,Hofmeister series ,Peptides/chemical synthesis ,TDP-43 ,Kinetics ,Liquid-Liquid Extraction ,Static Electricity ,lcsh:QR1-502 ,Cooperativity ,Biochemistry ,Article ,lcsh:Microbiology ,Hydrophobic effect ,03 medical and health sciences ,0302 clinical medicine ,Protein Domains ,Phase (matter) ,Liquid liquid ,Humans ,RNA/chemistry ,Recombinant Proteins/biosynthesis ,Amyotrophic Lateral Sclerosis/metabolism ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Science & Technology ,Chemistry ,aggregation ,protein aggregates ,Recombinant Proteins ,DNA-Binding Proteins ,Biophysics ,RNA ,Seeding ,AFM ,phase separation ,Peptides ,Life Sciences & Biomedicine ,Hydrophobic and Hydrophilic Interactions ,030217 neurology & neurosurgery ,DNA-Binding Proteins/chemistry ,Fluorescence Recovery After Photobleaching - Abstract
Aggregates of TAR DNA-binding protein (TDP-43) are a hallmark of several neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS). Although TDP-43 aggregates are an undisputed pathological species at the end stage of these diseases, the molecular changes underlying the initiation of aggregation are not fully understood. The aim of this study was to investigate how phase separation affects self-aggregation and aggregation seeded by pre-formed aggregates of either the low-complexity domain (LCD) or its short aggregation-promoting regions (APRs). By systematically varying the physicochemical conditions, we observed that liquid-liquid phase separation (LLPS) promotes spontaneous aggregation. However, we noticed less efficient seeded aggregation in phase separating conditions. By analyzing a broad range of conditions using the Hofmeister series of buffers, we confirmed that stabilizing hydrophobic interactions prevail over destabilizing electrostatic forces. RNA affected the cooperativity between LLPS and aggregation in a "reentrant" fashion, having the strongest positive effect at intermediate concentrations. Altogether, we conclude that conditions which favor LLPS enhance the subsequent aggregation of the TDP-43 LCD with complex dependence, but also negatively affect seeding kinetics. ispartof: BIOMOLECULES vol:11 issue:4 ispartof: location:Switzerland status: published
- Published
- 2021
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