Preparation of Amyloid Fibrils Seeded from Brain and Meninges

Scherpelz, Kathryn P; Lü, Junxia; Tycko, Robert; Meredith, Stephen C.

doi:10.1007/978-1-4939-2978-8_20

Cited by 11 publications

(18 citation statements)

References 20 publications

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“…For example, α-synuclein aggregates isolated from different types of synucleinopathies (e.g., PD and multiple system atrophy) exhibited drastically different structure (52,53) and seeding capacity, and induced different patterns of pathology spreading in in vitro neuronal cell models and animal models of synucleinopathies (16). Similar findings have been made for Aβ aggregates from AD brains and other amyloidogenic proteins (20,(54)(55)(56). While these differences have been attributed to differences in the structural properties of the fibrils present in these samples, direct visualization and characterization of some of the tissue-derived aggregates has not been possible.…”

Section: Discussionsupporting

confidence: 62%

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Cendrowska

Silva

Ait-Bouziad

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Increasing evidence suggests that amyloid polymorphism gives rise to different strains of amyloids with distinct toxicities and pathology-spreading properties. Validating this hypothesis is challenging due to a lack of tools and methods that allow for the direct characterization of amyloid polymorphism in hydrated and complex biological samples. Here, we report on the development of 11-mercapto-1-undecanesulfonate-coated gold nanoparticles (NPs) that efficiently label the edges of synthetic, recombinant, and native amyloid fibrils derived from different amyloidogenic proteins. We demonstrate that these NPs represent powerful tools for assessing amyloid morphological polymorphism, using cryogenic transmission electron microscopy (cryo-EM). The NPs allowed for the visualization of morphological features that are not directly observed using standard imaging techniques, including transmission electron microscopy with use of the negative stain or cryo-EM imaging. The use of these NPs to label native paired helical filaments (PHFs) from the postmortem brain of a patient with Alzheimer’s disease, as well as amyloid fibrils extracted from the heart tissue of a patient suffering from systemic amyloid light-chain amyloidosis, revealed a high degree of homogeneity across the fibrils derived from human tissue in comparison with fibrils aggregated in vitro. These findings are consistent with, and strongly support, the emerging view that the physiologic milieu is a key determinant of amyloid fibril strains. Together, these advances should not only facilitate the profiling and characterization of amyloids for structural studies by cryo-EM, but also pave the way to elucidate the structural basis of amyloid strains and toxicity, and possibly the correlation between the pathological and clinical heterogeneity of amyloid diseases.

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Section: Discussionsupporting

confidence: 62%

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Cendrowska

Silva

Ait-Bouziad

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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“…The "seeding" of amyloids is a process that represents triggering of the amyloid fibril formation by inoculation with preformed amyloid fibrils of the same or other proteins called "seeds." This process is mostly sequence-specific, and de novo forming fibrils exhibit close structural similarity or identity to the seeds [48,49].…”

Section: Amyloid Fibril Formation Of the Full-length Vicilin Can Be Ementioning

confidence: 99%

Accumulation of storage proteins in plant seeds is mediated by amyloid formation

et al. 2020

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Amyloids are protein aggregates with a highly ordered spatial structure giving them unique physicochemical properties. Different amyloids not only participate in the development of numerous incurable diseases but control vital functions in archaea, bacteria and eukarya. Plants are a poorly studied systematic group in the field of amyloid biology. Amyloid properties have not yet been demonstrated for plant proteins under native conditions in vivo. Here we show that seeds of garden pea Pisum sativum L. contain amyloid-like aggregates of storage proteins, the most abundant one, 7S globulin Vicilin, forms bona fide amyloids in vivo and in vitro. Full-length Vicilin contains 2 evolutionary conserved β-barrel domains, Cupin-1.1 and Cupin-1.2, that self-assemble in vitro into amyloid fibrils with similar physicochemical properties. However, Cupin-1.2 fibrils unlike Cupin-1.1 can seed Vicilin fibrillation. In vivo, Vicilin forms amyloids in the cotyledon cells that bind amyloid-specific dyes and possess resistance to detergents and proteases. The Vicilin amyloid accumulation increases during seed maturation and wanes at germination. Amyloids of Vicilin resist digestion by gastrointestinal enzymes, persist in canned peas, and exhibit toxicity for yeast and mammalian cells. Our finding for the first time reveals involvement of amyloid formation in the accumulation of storage proteins in plant seeds.

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“…These polymorphic structures are strongly influenced by the conditions of aggregation, including temperature, pH, metal ions, and physical agitation of the solution. Although little is known about the structure of fibril precursors, such as soluble oligomers, most of this polymorphism arises during nucleation, as is shown by the fact that seeding, which bypasses nucleation steps, leads to the formation of replicate progeny fibrils 1 – 3 , 31 , 32 .…”

Section: Introductionmentioning

confidence: 99%

Atomic-level differences between brain parenchymal- and cerebrovascular-seeded Aβ fibrils

Scherpelz

Wang

Pytel

et al. 2021

Sci Rep

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Alzheimer’s disease is characterized by neuritic plaques, the main protein components of which are β-amyloid (Aβ) peptides deposited as β-sheet-rich amyloid fibrils. Cerebral Amyloid Angiopathy (CAA) consists of cerebrovascular deposits of Aβ peptides; it usually accompanies Alzheimer’s disease, though it sometimes occurs in the absence of neuritic plaques, as AD also occurs without accompanying CAA. Although neuritic plaques and vascular deposits have similar protein compositions, one of the characteristic features of amyloids is polymorphism, i.e., the ability of a single pure peptide to adopt multiple conformations in fibrils, depending on fibrillization conditions. For this reason, we asked whether the Aβ fibrils in neuritic plaques differed structurally from those in cerebral blood vessels. To address this question, we used seeding techniques, starting with amyloid-enriched material from either brain parenchyma or cerebral blood vessels (using meninges as the source). These amyloid-enriched preparations were then added to fresh, disaggregated solutions of Aβ to make replicate fibrils, as described elsewhere. Such fibrils were then studied by solid-state NMR, fiber X-ray diffraction, and other biophysical techniques. We observed chemical shift differences between parenchymal vs. vascular-seeded replicate fibrils in select sites (in particular, Ala2, Phe4, Val12, and Gln15 side chains) in two-dimensional 13C-13C correlation solid-state NMR spectra, strongly indicating structural differences at these sites. X-ray diffraction studies also indicated that vascular-seeded fibrils displayed greater order than parenchyma-seeded fibrils in the “side-chain dimension” (~ 10 Å reflection), though the “hydrogen-bond dimensions” (~ 5 Å reflection) were alike. These results indicate that the different nucleation conditions at two sites in the brain, parenchyma and blood vessels, affect the fibril products that get formed at each site, possibly leading to distinct pathophysiological outcomes.

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Preparation of Amyloid Fibrils Seeded from Brain and Meninges

Cited by 11 publications

References 20 publications

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Accumulation of storage proteins in plant seeds is mediated by amyloid formation

Atomic-level differences between brain parenchymal- and cerebrovascular-seeded Aβ fibrils

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