9Y0S

CryoEM structure of alpha-synuclein fibril induced by psychosine

Summary for 9Y0S

• Entry DOI: 10.2210/pdb9y0s/pdb
• EMDB information: 72396
• Descriptor: Alpha-synuclein (1 entity in total)
• Functional Keywords: alpha-synuclein, amyloid, lipid, protein fibril
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 10
• Total formula weight: 144761.08

Authors

Jenkins, R.A.,Sun, C.Q.,Sawaya, M.R.,Rodriguez, J.A. (deposition date: 2025-08-29, release date: 2025-10-08)

Primary citation

Jenkins, R.A.,Wu, S.,Fujimura, G.,Heredia, A.,Flowers, C.W.,Sun, C.,Sawaya, M.R.,Loo, J.A.,Rodriguez, J.A.
Leveraging bioorthogonal conjugation for alpha synuclein fibril surveillance.
Biorxiv, 2025

PubMed Abstract: Alpha synuclein (α-syn) amyloid fibrils are associated with various neurodegenerative diseases. To better understand the molecular and cellular basis for α-syn fibril persistence and spread, we implemented a fluorophore labeling strategy to surveil pre-formed α-syn fibrils in solution and in cells. We leveraged amber codon mediated incorporation of a tetrazine-based artificial amino acid (TetV2.0) to install a cyclooctene-conjugated Janeliaflour, JF549, at four sites on human α-syn: residues 4, 60, 96 and 136. Fast coupling occurred under mild buffer conditions and in the presence of the disease-associated cofactor and cytotoxic lipid, psychosine. Labeled fibrils retained their polymorphic features, seeded the growth of new fibrils , and induced the seeding of positive puncta in α-syn FRET biosensor HEK293T cells. This allowed simultaneous tracking of exogenous and endogenous α-syn aggregates in biosensor cells, and their localization within the cells. In doing so, our approach facilitates more detailed mechanistic investigation of α-syn aggregates.

PubMed: 41000844
DOI: 10.1101/2025.09.12.675751

Experimental method

ELECTRON MICROSCOPY (2.72 Å)