An integrated Rheo-NMR platform for real-time, site-specific kinetic analysis of amyloid formation
Year: 2026
Type: preprint
Abstract: Amyloid fibrils are protein aggregates associated with neurodegenerative diseases. Their formation proceeds through protein– and condition-dependent microscopic pathways that can generate structurally and biologically distinct aggregates, yet these pathways are inferred from ensemble-averaged measurements. Amyloid formation is often slow under quiescent conditions, prompting agitation. However, conventional agitation precludes simultaneous high-resolution solution nuclear magnetic resonance (NMR) spectroscopy, limiting site-specific kinetic analysis of microscopic aggregation pathways. Here we present an integrated Rheo-NMR platform combining controlled shear flow, time-resolved NMR spectroscopy and mechanistic kinetic modeling to quantify site-specific amyloid-formation kinetics in a single, continuously monitored reaction. Applied to α-synuclein under dilute and polyethylene glycol (PEG)-induced liquid-liquid phase separation (LLPS) conditions, the method revealed environment-dependent redistribution of kinetic signatures associated with primary nucleation-elongation and secondary nucleation-elongation pathways across the sequence. The platform distinguishes changes in overall aggregation rates from differences in residue-specific kinetic profiles, providing a quantitative framework for dissecting environment-dependent amyloid formation.
Language: English
Subfield: Cellular and Molecular Neuroscience
Field: Neuroscience
Domain: Life Sciences
Open Access status: green
Awards: Rheo-NMR and paramagnetic relaxation-based structural determination of alpha-synuclein oligomers, Elucidation of the mechanism of amyloid fibrillation using Rheo-NMR and development of a new Rheo-NMR instrument, G3732707491, Velocity imaging of turbulence flow using Rheo-NMR, G6089999360, Elucidation of the mechanism of amyloid fibrillation of alpha-synuclein starting from liquid-liquid phase separation, G7082510209, G7622373350, G8433934829, Detection of amyloid polymorphism formation and high-resolution analysis
OpenAlex ID: W7223008630