An integrated Rheo-NMR platform for real-time, site-specific kinetic analysis of amyloid formation
- 1Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
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 -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.
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BibTeX
@misc{so2026integrated,
title = {{An integrated Rheo-NMR platform for real-time, site-specific kinetic analysis of amyloid formation}},
author = {M. So and K. Sato and K. Yoshioka and Y. Shimada and D. Morimoto and H. Nakamura and A. Furukawa and K. Sugase},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.19.760037},
url = {https://www.biorxiv.org/content/10.64898/2026.10.19.760037v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - An integrated Rheo-NMR platform for real-time, site-specific kinetic analysis of amyloid formation
AU - M. So
AU - K. Sato
AU - K. Yoshioka
AU - Y. Shimada
AU - D. Morimoto
AU - H. Nakamura
AU - A. Furukawa
AU - K. Sugase
PY - 2026
DA - 2026/10/19
PB - bioRxiv
DO - 10.64898/2026.10.19.760037
UR - https://www.biorxiv.org/content/10.64898/2026.10.19.760037v1
AB - 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 -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.
ER -