2.7 Å cryo-EM structure of ex vivo RML prion fibrils

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: July 13, 2022

Mammalian prions propagate as distinct strains and are composed of multichain assemblies misfolded host-encoded prion protein (PrP). Here, we present a near-atomic resolution cryo-EM structure PrP fibrils in highly infectious rod preparations isolated from the brains RML prion-infected mice. We found that rods comprise single-protofilament helical amyloid coexist with twisted pairs same protofilaments. Each rung protofilament is formed by single monomer ordered core comprising residues 94-225, which folds to create two asymmetric lobes N-linked glycans glycosylphosphatidylinositol anchor projecting C-terminal lobe. The overall architecture comparable recently reported brain hamsters infected 263K strain. However, there marked conformational variations could result differences sequence and/or represent distinguishing features strains.

Language: Английский

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Cryo-EM structure of anchorless RML prion reveals variations in shared motifs between distinct strains

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: July 13, 2022

Little is known about the structural basis of prion strains. Here we provide a high (3.0 Å) resolution cryo-electron microscopy-based structure infectious brain-derived fibrils mouse anchorless RML scrapie strain which, like recently determined hamster 263K strain, has parallel in-register β-sheet-based core. Several motifs are shared between these ex vivo strains, including an amino-proximal steric zipper and three β-arches. However, detailed comparisons reveal variations in topologies other features. Unlike wildtype prions, prions lack glycophosphatidylinositol anchors severely deficient N-linked glycans. Nonetheless, similarity our to one reported for accompanying paper indicates that post-translational modifications do not substantially alter amyloid core conformation. This work demonstrates both common divergent features strains at near-atomic level.

Language: Английский

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A structural basis for prion strain diversity

Nature Chemical Biology, Journal Year: 2023, Volume and Issue: 19(5), P. 607 - 613

Published: Jan. 16, 2023

Abstract Recent cryogenic electron microscopy (cryo-EM) studies of infectious, ex vivo, prion fibrils from hamster 263K and mouse RML strains revealed a similar, parallel in-register intermolecular β-sheet (PIRIBS) amyloid architecture. Rungs the are composed individual protein (PrP) monomers that fold to create distinct N-terminal C-terminal lobes. However, disparity in hamster/mouse PrP sequence precludes understanding how divergent emerge an identical substrate. In this study, we determined near-atomic resolution cryo-EM structure vivo ME7 strain compared with fibril structure. This structural comparison two biologically mouse-adapted suggests defined folding subdomains rungs way which they interrelated, providing definition intra-species strain-specific conformations.

Language: Английский

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Misfolded protein oligomers: mechanisms of formation, cytotoxic effects, and pharmacological approaches against protein misfolding diseases

Molecular Neurodegeneration, Journal Year: 2024, Volume and Issue: 19(1)

Published: Feb. 20, 2024

The conversion of native peptides and proteins into amyloid aggregates is a hallmark over 50 human disorders, including Alzheimer's Parkinson's diseases. Increasing evidence implicates misfolded protein oligomers produced during the formation process as primary cytotoxic agents in many these devastating conditions. In this review, we analyze processes by which are formed, their structures, physicochemical properties, population dynamics, mechanisms cytotoxicity. We then focus on drug discovery strategies that target ability to disrupt cell physiology trigger degenerative processes.

Language: Английский

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Creutzfeldt–Jakob disease and other prion diseases

Nature Reviews Disease Primers, Journal Year: 2024, Volume and Issue: 10(1)

Published: Feb. 29, 2024

Language: Английский

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Cryo-EM structures of prion protein filaments from Gerstmann–Sträussler–Scheinker disease

Acta Neuropathologica, Journal Year: 2022, Volume and Issue: 144(3), P. 509 - 520

Published: July 12, 2022

Prion protein (PrP) aggregation and formation of PrP amyloid (APrP) are central events in the pathogenesis prion diseases. In dominantly inherited amyloidosis known as Gerstmann-Sträussler-Scheinker (GSS) disease, plaques made present throughout brain. The c.593t > c mutation gene (PRNP) results a phenylalanine to serine amino acid substitution at residue 198 (F198S) causes most severe among GSS variants. It has been shown that neurodegeneration this disease is associated with presence extracellular APrP neuronal intracytoplasmic Tau inclusions, have contain paired helical filaments identical those found Alzheimer disease. Using cryogenic electron microscopy (cryo-EM), we determined for first time structures human APrP, isolated post-mortem from brain two symptomatic PRNP F198S carriers. We report composed dimeric, trimeric tetrameric left-handed protofilaments their protomers sharing common fold. cross-β spines consist 62 acids span glycine 80 141, adopting previously unseen spiral fold thicker outer layer thinner inner layer. Each protomer comprises nine short β-strands, β1 β8 strands, well β4 β9 forming steric zipper. data obtained by cryo-EM provide insights into structural complexity filament amyloidosis. novel findings highlight urgency extending our knowledge filaments' may underlie distinct clinical pathologic phenotypes neurodegenerative

Language: Английский

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High-throughput cryo-EM structure determination of amyloids

Faraday Discussions, Journal Year: 2022, Volume and Issue: 240, P. 243 - 260

Published: Jan. 1, 2022

This paper presents new cryo-EM image processing methods for amyloids, including automated picking and strategies helical structure determination in RELION. The shows examples of tau filament structures that were solved using these methods.

Language: Английский

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Ultrastructural and biochemical classification of pathogenic tau, α-synuclein and TDP-43

Acta Neuropathologica, Journal Year: 2022, Volume and Issue: 143(6), P. 613 - 640

Published: May 5, 2022

Abstract Intracellular accumulation of abnormal proteins with conformational changes is the defining neuropathological feature neurodegenerative diseases. The pathogenic that accumulate in patients' brains adopt an amyloid-like fibrous structure and exhibit various ultrastructural features. biochemical analysis sarkosyl-insoluble fractions extracted from patients’ also shows disease-specific Intriguingly, these features are common within same disease group. These differences among have important implications for definitive diagnosis disease, suggest existence protein strains contribute to heterogeneity pathogenesis Recent experimental evidence has shown prion-like propagation host cells recipient underlies onset progression reproduction pathological characterize each cellular animal models implies structural inherited a manner. In this review, we summarize patients diseases forms tau, α-synuclein, TDP-43, discuss how properties maintained brain, based on recent insights.

Language: Английский

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Cryo-EM of prion strains from the same genotype of host identifies conformational determinants

PLoS Pathogens, Journal Year: 2022, Volume and Issue: 18(11), P. e1010947 - e1010947

Published: Nov. 7, 2022

Prion strains in a given type of mammalian host are distinguished by differences clinical presentation, neuropathological lesions, survival time, and characteristics the infecting prion protein (PrP) assemblies. Near-atomic structures prions from two species with different PrP sequences have been determined but comparisons distinct same amino acid sequence needed to identify purely conformational determinants strain characteristics. Here we report 3.2 Å resolution cryogenic electron microscopy-based structure 22L purified brains mice engineered express only lacking glycophosphatidylinositol anchors [anchorless (a) 22L]. Comparison this near-atomic our recently aRML propagated inbred mouse reveals that these templates for growth via incorporation molecules sequence. Both a22L assembled as stacks forming parallel in-register intermolecular β-sheets intervening loops, single monomers spanning ordered fibril core. Each monomer shares an N-terminal steric zipper, three major arches, overall V-shape, details other features differ markedly. Thus, variations shared motifs within β-stack architecture provide structural basis differentiation genotype.

Language: Английский

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The brain interactome of a permissive prion replication substrate

Neurobiology of Disease, Journal Year: 2025, Volume and Issue: unknown, P. 106802 - 106802

Published: Jan. 1, 2025

Bank voles are susceptible to prion strains from many different species, yet the molecular mechanisms underlying ability of bank vole protein (BVPrP) function as a universal acceptor remain unclear. Potential differences in environments and interaction networks on cell surface brain cells may contribute BVPrP's unusual behavior. To test this hypothesis, we generated knock-in mice that express physiological levels BVPrP (M109 isoform) employed mass spectrometry compare interactomes mouse (Mo) PrP following mild vivo crosslinking tissue. Substantial overlap was observed between top interactors for MoPrP, with established PrP-interactors such neural adhesion molecules, subunits Na

Language: Английский

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