Inertial Agitation in an NMR Magnet: Real-Time Monitoring of Protein Aggregation at High Resolution.

Area of Science:

• Biochemistry
• Neuroscience
• Spectroscopy

Background:

• Protein aggregation is implicated in various neurodegenerative diseases.
• Monitoring protein aggregation in real-time is crucial for understanding disease mechanisms.
• Current methods for inducing and monitoring aggregation can be limited.

Purpose of the Study:

• To develop a novel method for inducing and monitoring protein aggregation inside an NMR magnet.
• To investigate the role of protein interactions in aggregation pathways.
• To explore the diagnostic potential of this method for neurodegenerative diseases.

Main Methods:

• Developed "inertia" NMR, utilizing rotational inertia of an eccentric floater for sample agitation.
• Combined inertia NMR with 2D NMR for high-resolution aggregation studies.
• Employed isotope editing/filtering for tracking protein coaggregation.

Main Results:

• Successfully induced and monitored aggregation of disease-associated proteins under near-physiological conditions.
• Observed accelerated aggregation of alpha-synuclein (αS) by its splicing isoform and TDP-43 CTD.
• Demonstrated diagnostic potential by differentiating Parkinson's disease from MSA using patient-derived seeds.

Conclusions:

• Inertia NMR provides high-resolution insights into protein aggregation dynamics.
• The method reveals cross-seeding roles in synucleinopathies.
• This technique offers a general strategy for accelerating chemical processes via efficient mixing in NMR.