Combining light-induced aggregation and biotin proximity labeling implicates endolysosomal proteins in early...

Area of Science:

• Neuroscience
• Molecular Biology
• Biochemistry

Background:

• Alpha-synuclein (α-syn) aggregation is a hallmark of Parkinson's disease (PD) and related synucleinopathies.
• Understanding the early stages of α-syn aggregation is critical but challenging due to limitations in current experimental tools.

Purpose of the Study:

• To introduce and validate UltraID-LIPA, a novel platform for identifying α-syn-interacting proteins.
• To uncover molecular mechanisms driving α-syn oligomerization in the early stages of aggregation.

Main Methods:

• Integration of the light-inducible protein aggregation (LIPA) system with the UltraID proximity-dependent biotinylation assay.
• Application of the UltraID-LIPA platform to identify proteins interacting with α-syn during aggregation.

Main Results:

• Identification of 38 α-syn-interacting proteins, including known and novel candidates.
• Demonstration of the platform's accuracy and robustness in capturing dynamic protein interactions.
• Observation of significant interactions with endolysosomal and membrane-associated proteins, implicating organelles in early aggregation.

Conclusions:

• UltraID-LIPA is a powerful tool for studying dynamic protein aggregation events.
• Early α-syn aggregation is strongly linked to interactions with membrane-bound organelles.
• The findings provide new insights into the pathogenesis of synucleinopathies and other proteinopathies.