Protein Misfolding Cyclic Amplification of Prions

Saunders, Samuel E.; Bartz, Jason C.; Shikiya, Ronald A.

doi:10.3791/4075

Cited by 5 publications

(3 citation statements)

References 25 publications

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“…Or, as more prohibitively stated: “Prions are composed solely of the disease-causing prion protein (PrP Sc )” 7 . This exclusive view continues to be promulgated 8 , 9 even though other molecules, including nucleic acids, can be required for replication of TSE infectious agents 10 …”

Section: Introductionmentioning

confidence: 99%

Infectious particles, stress, and induced prion amyloids

Manuelidis

2013

Virulence

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Transmissible encephalopathies (TSEs) are believed by many to arise by spontaneous conversion of host prion protein (PrP) into an infectious amyloid (PrP-res, PrPSc) without nucleic acid. Many TSE agents reside in the environment, with infection controlled by public health measures. These include the disappearance of kuru with the cessation of ritual cannibalism, the dramatic reduction of epidemic bovine encephalopathy (BSE) by removal of contaminated feed, and the lack of endemic scrapie in geographically isolated Australian sheep with susceptible PrP genotypes. While prion protein modeling has engendered an intense focus on common types of protein misfolding and amyloid formation in diverse organisms and diseases, the biological characteristics of infectious TSE agents, and their recognition by the host as foreign entities, raises several fundamental new directions for fruitful investigation such as: (1) unrecognized microbial agents in the environmental metagenome that may cause latent neurodegenerative disease, (2) the evolutionary social and protective functions of different amyloid proteins in diverse organisms from bacteria to mammals, and (3) amyloid formation as a beneficial innate immune response to stress (infectious and non-infectious). This innate process however, once initiated, can become unstoppable in accelerated neuronal aging.

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Section: Introductionmentioning

confidence: 99%

Infectious particles, stress, and induced prion amyloids

Manuelidis

2013

Virulence

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“…PrP Sc generated through PMCA shares similar structural and biochemical properties as PrP Sc generated in an animal bioassay and is infectious in animal bioassays [ 20 , 21 , 22 , 23 ]. PMCA can be used to analyze low titre samples, examine species-barrier effects and evaluate the role of potential cofactors (polyanionic components, lipids and proteoglycans [ 1 , 24 , 25 , 26 ]) involved in PrP Sc generation [ 21 , 27 , 28 ]. PMCA analysis of samples can, however, be confounded by the de nova generation of PrP Res via off-target amplification of normal non-infectious material [ 29 ].…”

Section: Introductionmentioning

confidence: 99%

The Standard Scrapie Cell Assay: Development, Utility and Prospects

Merwe

Aiken

Westaway

et al. 2015

Viruses

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Prion diseases are a family of fatal neurodegenerative diseases that involve the misfolding of a host protein, PrPC. Measuring prion infectivity is necessary for determining efficacy of a treatment or infectivity of a prion purification procedure; animal bioassays are, however, very expensive and time consuming. The Standard Scrapie Cell Assay (SSCA) provides an alternative approach. The SSCA facilitates quantitative in vitro analysis of prion strains, titres and biological properties. Given its robust nature and potential for high throughput, the SSCA has substantial utility for in vitro characterization of prions and can be deployed in a number of settings. Here we provide an overview on establishing the SSCA, its use in studies of disease dissemination and pathogenesis, potential pitfalls and a number of remaining challenges.

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“…65 Briefly, we used a Misonix 3000 sonicator (Farmingdale, NY) coupled to a 96-well plate titanium cup-horn. The sonicator output was set to level 6.…”

Section: Prion Strains and Negative Controlsmentioning

confidence: 99%

Prion formation, but not clearance, is supported by protein misfolding cyclic amplification

Shikiya

Eckland

Young

et al. 2014

Prion

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Prion diseases are fatal transmissible neurodegenerative disorders that affect animals including humans. The kinetics of prion infectivity and PrP Sc accumulation can differ between prion strains and within a single strain in different tissues. The net accumulation of PrP Sc in animals is controlled by the relationship between the rate of PrP Sc formation and clearance. Protein misfolding cyclic amplification (PMCA) is a powerful technique that faithfully recapitulates PrP Sc formation and prion infectivity in a cell-free system. PMCA has been used as a surrogate for animal bioassay and can model species barriers, host range, strain co-factors and strain interference. In this study we investigated if degradation of PrP Sc and/or prion infectivity occurs during PMCA. To accomplish this we performed PMCA under conditions that do not support PrP Sc formation and did not observe either a reduction in PrP Sc abundance or an extension of prion incubation period, compared to untreated control samples. These results indicate that prion clearance does not occur during PMCA. These data have significant implications for the interpretation of PMCA based experiments such as prion amplification rate, adaptation to new species and strain interference where production and clearance of prions can affect the outcome.

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Protein Misfolding Cyclic Amplification of Prions

Cited by 5 publications

References 25 publications

Infectious particles, stress, and induced prion amyloids

Infectious particles, stress, and induced prion amyloids

The Standard Scrapie Cell Assay: Development, Utility and Prospects

Prion formation, but not clearance, is supported by protein misfolding cyclic amplification

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