The physical dimensions of amyloid aggregates control their infective potential as prion particles

Marchante, Ricardo; Béal, David; Koloteva-Levine, Nadejda; Purton, Tracey J; Tuite, Mick F.; Xue, Wei‐Feng

doi:10.7554/elife.27109

Cited by 36 publications

(51 citation statements)

References 51 publications

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“…A trivial explanation for this observation is that the change in Sup35p assemblies size (Fig. B) affects their ability to reach the cytosol of recipient [ psi ‐ ] cells where they convert Sup35p monomers to the prion conformation (Marchante et al , ). The infectivity of [ PSI + ] cell lysates slowly declined after 24 h of growth (Fig.…”

Section: Resultsmentioning

confidence: 99%

Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI⁺] prion in yeast

Wang¹,

Melki

Kabani

2019

Molecular Microbiology

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Summary The translation termination factor Sup35p can form self‐replicating fibrillar aggregates responsible for the [PSI+] prion state. Sup35p aggregation yields detergent‐resistant assemblies detectable on agarose gels under semi‐denaturant conditions and fluorescent puncta within the yeast cytosol when the protein is fused to GFP. It is still unclear whether any of these manifestations of [PSI+] truly correspond to the Sup35p assemblies that faithfully transmit the [PSI+] prion from mother to daughter cells. The infectious titer of prions in cells can be indirectly assessed by the ability of [PSI+] cells lysates to induce the prion state when introduced into naïve cells. Here, we report that the dramatic changes in the size and amounts of SDS‐resistant Sup35p that occur during growth do not correlate with the infectious titer. Our results suggest that fluorescent Sup35‐GFP puncta and detergent‐resistant Sup35p assemblies are good indicators of Sup35p conversion to the prion state but not of infectious particles number.

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Section: Resultsmentioning

confidence: 99%

Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI⁺] prion in yeast

Wang¹,

Melki

Kabani

2019

Molecular Microbiology

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“…The peptide samples were dissolved in 6M GdnHCl at pH10 and Aβ42 monomers were purified by gel filtration using a Superdex 75 column immediately prior to assembly in order to ensure the generation of reproducible and high quality Aβ42 amyloid seeds (Hellstrand et al, 2010;Walsh et al, 2009). Monomeric Sup35NM protein was produced recombinantly in E. coli and assembled as described previously (Marchante et al, 2017). Aβ42 and Sup35NM fibrils were subsequently dispersed by brief controlled sonication (see Materials and Methods) and imaged using atomic force microscopy (AFM).…”

Section: Self-seeded Growth Of Both Aβ42 and Sup35nm Amyloid Fibrils mentioning

confidence: 99%

Amyloid particles facilitate surface-catalyzed cross-seeding by acting as promiscuous nanoparticles

Koloteva-Levine

Marchante

Purton

et al. 2020

Preprint

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Amyloid seeds are nanometre-sized protein particles that accelerate amyloid assembly, as well as propagate and transmit the amyloid protein conformation associated with a wide range of protein misfolding diseases. However, seeded amyloid growth through templated elongation at fibril ends cannot explain the full range of molecular behaviours observed during cross-seeded formation of amyloid by heterologous seeds. Here, we demonstrate that amyloid seeds can accelerate amyloid formation via a surface catalysis mechanism without propagating the specific amyloid conformation associated with the seeds. This type of seeding mechanism is demonstrated through quantitative characterisation of the cross-seeded assembly reactions involving two non-homologous and unrelated proteins: the human Aβ42 peptide and the yeast prion-forming protein Sup35NM. Our results suggest experimental approaches to differentiate seeding by templated elongation from non-templated amyloid seeding, and rationalise the molecular mechanism of the cross-seeding phenomenon as a manifestation of the aberrant surface activities presented by amyloid seeds as nanoparticles.

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“…Thus, fibril polymorphs displaying different properties would likely 343 have different biological activities, even formed from identical precursors and present within 344 the same population. In this case, individual fibrils within a population may also have higher 345 rates of fragmentation that could result in a greater likelihood for propagation 51,52 or 346 cytotoxicity 34 , or higher rates of secondary nucleation that could result in more cytotoxic active 347…”

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confidence: 99%

Quantification of amyloid fibril polymorphism by nano-morphometry reveals the individuality of filament assembly

Aubrey

Blakeman

Lutter

et al. 2020

Preprint

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Amyloid fibrils are highly polymorphic structures formed by many different proteins. They provide biological function but also abnormally accumulate in numerous human diseases. The physicochemical principles of amyloid polymorphism are not understood due to lack of structural insights at the single-fibril level. To identify and classify different fibril polymorphs and to quantify the level of heterogeneity is essential to decipher the precise links between amyloid structures and their functional and disease associated properties such as toxicity, strains, propagation and spreading. Employing gentle, force-curve based AFM, we produce detailed images, from which the 3D reconstruction of individual filaments in heterogeneous amyloid samples is achieved. Distinctive fibril polymorphs are then classified by hierarchical clustering, and sample heterogeneity is objectively quantified. These data demonstrate the polymorphic nature of fibril populations, provide important information regarding the energy landscape of amyloid self-assembly, and offer quantitative insights into the structural basis of polymorphism in amyloid populations.

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The physical dimensions of amyloid aggregates control their infective potential as prion particles

Cited by 36 publications

References 51 publications

Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI⁺] prion in yeast

Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI⁺] prion in yeast

Amyloid particles facilitate surface-catalyzed cross-seeding by acting as promiscuous nanoparticles

Quantification of amyloid fibril polymorphism by nano-morphometry reveals the individuality of filament assembly

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The physical dimensions of amyloid aggregates control their infective potential as prion particles

Cited by 36 publications

References 51 publications

Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI+] prion in yeast

Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI+] prion in yeast

Amyloid particles facilitate surface-catalyzed cross-seeding by acting as promiscuous nanoparticles

Quantification of amyloid fibril polymorphism by nano-morphometry reveals the individuality of filament assembly

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Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI⁺] prion in yeast

Growth phase‐dependent changes in the size and infectivity of SDS‐resistant Sup35p assemblies associated with the [PSI⁺] prion in yeast