Distinct Stability States of Disease-Associated Human Prion Protein Identified by Conformation-Dependent Immunoassay

Choi, Young Pyo; Peden, Alexander; Gröner, Albrecht; Ironside, James W.; Head, Mark W.

doi:10.1128/jvi.01057-10

Cited by 27 publications

(29 citation statements)

References 53 publications

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“…In this study, the same atypical features were found to characterise PrP Sc from other sporadic human and animal TSEs co-analysed with GSS subtypes, well distinguishable from classical types of PrP Sc found in CJD and scrapie. Indeed, when analysed with similar approaches in previous studies, CJD and classical scrapie didn’t show evidence of being enriched in insoluble and protease sensitive PrP Sc [58], [69]. Furthermore, although PK cleavages other than those at residues 82 and 97 have been reported in sCJD [67], scrapie [70] and H-type BSE [71], in all these instances the glycosylated C-terminus of PrP Sc was always part of the protease resistant core.…”

Section: Discussionmentioning

confidence: 67%

Small Ruminant Nor98 Prions Share Biochemical Features with Human Gerstmann-Sträussler-Scheinker Disease and Variably Protease-Sensitive Prionopathy

et al. 2013

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Prion diseases are classically characterized by the accumulation of pathological prion protein (PrPSc) with the protease resistant C-terminal fragment (PrPres) of 27–30 kDa. However, in both humans and animals, prion diseases with atypical biochemical features, characterized by PK-resistant PrP internal fragments (PrPres) cleaved at both the N and C termini, have been described. In this study we performed a detailed comparison of the biochemical features of PrPSc from atypical prion diseases including human Gerstmann-Sträussler-Scheinker disease (GSS) and variably protease-sensitive prionopathy (VPSPr) and in small ruminant Nor98 or atypical scrapie. The kinetics of PrPres production and its cleavage sites after PK digestion were analyzed, along with the PrPSc conformational stability, using a new method able to characterize both protease-resistant and protease-sensitive PrPSc components. All these PrPSc types shared common and distinctive biochemical features compared to PrPSc from classical prion diseases such as sporadic Creutzfeldt-Jakob disease and scrapie. Notwithstanding, distinct biochemical signatures based on PrPres cleavage sites and PrPSc conformational stability were identified in GSS A117V, GSS F198S, GSS P102L and VPSPr, which allowed their specific identification. Importantly, the biochemical properties of PrPSc from Nor98 and GSS P102L largely overlapped, but were distinct from the other human prions investigated. Finally, our study paves the way towards more refined comparative approaches to the characterization of prions at the animal–human interface.

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Section: Discussionmentioning

confidence: 67%

Small Ruminant Nor98 Prions Share Biochemical Features with Human Gerstmann-Sträussler-Scheinker Disease and Variably Protease-Sensitive Prionopathy

et al. 2013

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“…This method allowed us to compare the conformational features of human PrP Sc independently of proteolytic treatment and in addition provided quantitative data on levels of PrP Sc , sPrP Sc , and rPrP Sc [15], [30]. The CDI techniques represent a major improvement over previously used semi-quantitative WB-based methods, the finding that has been independently confirmed by another group [60], [61]. The dissociation and unfolding of PrPSc in a presence of increasing concentration of Gdn HCl can be described as follows: [PrP Sc ] n →[sPrP Sc ] n →iPrP→uPrP, where [PrP Sc ] n are native aggregates of PrP Sc , [sPrP Sc ] n are soluble protease-sensitive oligomers of PrP Sc , iPrP is an intermedite, and uPrP is completely unfolded (denatured) PrP [7], [43], [62].…”

Section: Discussionmentioning

confidence: 90%

Protease-Sensitive Conformers in Broad Spectrum of Distinct PrPSc Structures in Sporadic Creutzfeldt-Jakob Disease Are Indicator of Progression Rate

et al. 2011

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The origin, range, and structure of prions causing the most common human prion disease, sporadic Creutzfeldt-Jakob disease (sCJD), are largely unknown. To investigate the molecular mechanism responsible for the broad phenotypic variability of sCJD, we analyzed the conformational characteristics of protease-sensitive and protease-resistant fractions of the pathogenic prion protein (PrPSc) using novel conformational methods derived from a conformation-dependent immunoassay (CDI). In 46 brains of patients homozygous for polymorphisms in the PRNP gene and exhibiting either Type 1 or Type 2 western blot pattern of the PrPSc, we identified an extensive array of PrPSc structures that differ in protease sensitivity, display of critical domains, and conformational stability. Surprisingly, in sCJD cases homozygous for methionine or valine at codon 129 of the PRNP gene, the concentration and stability of protease-sensitive conformers of PrPSc correlated with progression rate of the disease. These data indicate that sCJD brains exhibit a wide spectrum of PrPSc structural states, and accordingly argue for a broad spectrum of prion strains coding for different phenotypes. The link between disease duration, levels, and stability of protease-sensitive conformers of PrPSc suggests that these conformers play an important role in the pathogenesis of sCJD.

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“…At variance with experimentally cloned rodent prion strains, however, less numerous and less conclusive data have been obtained with PrP Sc preparations extracted from naturally occurring prion diseases, especially in humans. In the few studies performed to date, sCJD MM1 PrP Sc was shown to be more stable than MM 2C PrP Sc by both CDI and CSA, and MM1 PrP Sc was shown to have a moderately higher stability than vCJD PrP Sc by CDI (32)(33)(34). In the present study, we have carried out the first systematic analysis of PrP Sc conformational stability in a large series of brain samples across the whole spectrum of human sCJD and vCJD prions.…”

Section: Discussionmentioning

confidence: 99%

“…In particular, a systematic analysis of the conformational stability of PrP Sc aggregates across the spectrum of human prions is still lacking. Previous studies focused on the comparison between sCJD subtypes MM1 and MM2 (31)(32)(33) or between variant CJD (vCJD) and the most common sCJD type MM1 (34). Furthermore, only the unfolding induced by GdnHCl was addressed, whereas the thermostability of PrP Sc has never been explored in CJD.…”

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confidence: 99%

Analysis of Conformational Stability of Abnormal Prion Protein Aggregates across the Spectrum of Creutzfeldt-Jakob Disease Prions

Cescatti

Saverioni

Capellari

et al. 2016

J Virol

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The wide phenotypic variability of prion diseases is thought to depend on the interaction of a host genotype with prion strains that have self-perpetuating biological properties enciphered in distinct conformations of the misfolded prion protein PrP Sc . This concept is largely based on indirect approaches studying the effect of proteases or denaturing agents on the physicochemical properties of PrP Sc aggregates. Furthermore, most data come from studies on rodent-adapted prion strains, making current understanding of the molecular basis of strains and phenotypic variability in naturally occurring diseases, especially in humans, more limited. To fill this gap, we studied the effects of guanidine hydrochloride (GdnHCl) and heating on PrP Sc aggregates extracted from 60 sporadic Creutzfeldt-Jakob disease (CJD) and 6 variant CJD brains. While denaturation curves obtained after exposure of PrP Sc to increasing GdnHCl concentrations showed similar profiles among the 7 CJD types analyzed, PrP Sc exposure to increasing temperature revealed significantly different and type-specific responses. In particular, MM1 and VV2, the most prevalent and fast-replicating CJD types, showed stable and highly resistant PrP Sc aggregates, whereas VV1, a rare and slowly propagating type, revealed unstable aggregates that easily dissolved at low temperature. Taken together, our results indicate that the molecular interactions mediating the aggregation state of PrP Sc , possibly enciphering strain diversity, are differently targeted by GdnHCl, temperature, and proteases. Furthermore, the detected positive correlation between the thermostability of PrP Sc aggregates and disease transmission efficiency makes inconsistent the proposed hypothesis that a decrease in conformational stability of prions results in an increase in their replication efficiency. IMPORTANCEPrion strains are defined as infectious isolates propagating distinctive phenotypic traits after transmission to syngeneic hosts. Although the molecular basis of prion strains is not fully understood, it is largely accepted that variations in prion protein conformation drive the molecular changes leading to the different phenotypes. In this study, we exposed abnormal prion protein aggregates encompassing the spectrum of human prion strains to both guanidine hydrochloride and thermal unfolding. Remarkably, while exposure to increasing temperature revealed significant strain-specific differences in the denaturation profile of the protein, treatment with guanidine hydrochloride did not. The findings suggest that thermal and chemical denaturation perturb the structure of prion protein aggregates differently. Moreover, since the most thermostable prion protein types were those associated with the most prevalent phenotypes and most rapidly and efficiently transmitting strains, the results suggest a direct correlation between strain replication efficiency and the thermostability of prion protein aggregates. P rion diseases are invariably fatal neurodegenerative disorders of humans ...

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Distinct Stability States of Disease-Associated Human Prion Protein Identified by Conformation-Dependent Immunoassay

Cited by 27 publications

References 53 publications

Small Ruminant Nor98 Prions Share Biochemical Features with Human Gerstmann-Sträussler-Scheinker Disease and Variably Protease-Sensitive Prionopathy

Small Ruminant Nor98 Prions Share Biochemical Features with Human Gerstmann-Sträussler-Scheinker Disease and Variably Protease-Sensitive Prionopathy

Protease-Sensitive Conformers in Broad Spectrum of Distinct PrPSc Structures in Sporadic Creutzfeldt-Jakob Disease Are Indicator of Progression Rate

Analysis of Conformational Stability of Abnormal Prion Protein Aggregates across the Spectrum of Creutzfeldt-Jakob Disease Prions

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