High-throughput cryo-EM structure determination of amyloids

Area of Science:

• Structural Biology
• Biophysics
• Neurodegenerative Diseases

Background:

• Amyloid filament formation is a hallmark of degenerative diseases.
• Electron cryo-microscopy (cryo-EM) has enabled atomic structure determination of amyloid filaments.
• Observed amyloid structures can differ between in vitro assembly and diseased tissues, complicating disease modeling.

Purpose of the Study:

• To identify factors governing the formation of distinct amyloid folds.
• To develop relevant model systems for studying degenerative diseases.
• To accelerate atomic structure determination of amyloids for high-throughput screening.

Main Methods:

• Development of a novel filament picking algorithm based on the Topaz approach.
• Implementation of optimized image processing strategies within Relion software.
• Application of high-throughput cryo-EM for rapid structure determination.

Main Results:

• A new algorithm and processing pipeline enable rapid atomic structure determination of amyloid filaments.
• The methods facilitate screening of numerous in vitro assembly conditions.
• Atomic structures of amyloids can now be determined within days.

Conclusions:

• The developed computational tools significantly enhance the speed and efficiency of amyloid structure determination.
• This advancement is crucial for understanding the molecular mechanisms underlying amyloid-related diseases.
• High-throughput cryo-EM, powered by these new algorithms, will facilitate the discovery of disease-specific amyloid structures.