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Cryo-EM study of alpha-synuclein fibrils with hereditary disease mutation


EMSL Project ID
51251

Abstract

With recent advances of cryo-EM technologies, including direct electron detecting cameras, we are now able to probe a broad scope of amyloid fibril structures to a level of molecular detail that has not been previously achieved. Our preliminary experiments determined cryo-EM structures of two alpha-synuclein fibril polymorphs with resolutions of 3.7 Å (Li et al., Nature Comminications 2018). Our structural determination efforts have been expanded to mutant alpha-synuclein fibrils. We aim to compare the structures of the mutant fibrils and the wild-type fibrils to assess how sequence changes of the disease-related mutations affect their structures and thus their biological activity. We have determined the structures of the two mutant fibrils (H50Q, E46K) grown under the same conditions that wild-type alpha-synuclein fibril structures are determined previously (Boyer et al., Nat. Struct. Mol. Bio. 2019; Boyer et al, under revision). We will also perform the cryo-EM study of other disease mutant alpha-synuclein fibrils. Structural comparison between the fibril seeds and resulting aggregates formed in cells will explore how a particular fibril conformation is preserved and amplified during the cellular seeding process. Our preliminary negative stain EM study of disease mutant alpha-synuclein fibrils demonstrate the feasibility and significance of our proposed approach, laying the groundwork to correlate structural differences to the biological activity changes in seeding and toxicity. This work will determine which fibril structure is responsible for the observed activity, allowing for a better understanding of structure-activity relationship of different disease mutant synuclein fibrils.

Project Details

Start Date
2020-01-15
End Date
2021-03-17
Status
Closed

Team

Principal Investigator

Lin Jiang
Institution
University of California, Los Angeles

Team Members

Binsen Li
Institution
University of California, Los Angeles

Nancy Meyer
Institution
Oregon Health & Science University

Irina El Khoury
Institution
Environmental Molecular Sciences Laboratory