Characterization of proteinase K-resistant N- and C-terminally truncated PrP in Nor98 atypical scrapie

Klingeborn, Mikael; Wik, Lotta; Simonsson, Magnus; Renström, Lena; Ottinger, Therese; Linné, Tommy

doi:10.1099/vir.0.81618-0

Cited by 56 publications

(63 citation statements)

References 45 publications

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“…Recently, three published biological studies have given a more precise description of newly identified fragments. The atypical/Nor98 scrapie isolates display after proteinase K treatment a distinct multiple band pattern, with a fast migrating band reported to be around 11-12 kDa [2, 5, 18] or 7-8 kDa [24,31] in size. These differences between the atypical/Nor98 scrapie and the classical scrapie isolates are illustrated in Figure 2. It is noteworthy that, in contrast to what has been reported in human TSE, the same multiple band Western blot profile is observed independently of the brain region [2,31].…”

Section: Identification Of New Prp Sc Fragmentsmentioning

confidence: 99%

“…Klingeborn et al [24] used harsher proteinase K conditions, which, together with the applied antibodies, might explain at least partially the differences between the two studies. One of the isolates of the Swedish isolate has been analysed with the method used by Arsac et al [2] (TeSeE Western blot, Bio-Rad), giving in these conditions a Western blot profile which was identical to the Norwegian Nor98 isolates.…”

Section: Deglycosylation Experimentsmentioning

confidence: 99%

“…Three recent studies have contributed to new information concerning the cleavage sites of the atypical/Nor98 scrapie PrP Sc [2,18,24]. (ii) Fragment B could be N-terminally (∼ aa 120) and C-terminally (∼ aa 233) cleaved.…”

Section: Prp Sc Cleavage Sitesmentioning

confidence: 99%

“…(iii) The smallest fragment, called fragment C by Arsac et al [2] and PrP7 by Klingeborn et al [24], could correspond to an N (∼ aa 85) and C (∼ aa 148-155) terminally truncated PrPres protein (∼ aa 90 and ∼ aa 153 for Gretzschel et al [18]) as estimated based on the mAb-binding sites retained. It is speculated that the truncation of the C-terminus might occur in vivo, since the C fragment is not detectable in non-proteinase K treated isolates when labelled with antibodies raised against the sequence situated upstream of aa 153 [18].…”

Section: Prp Sc Cleavage Sitesmentioning

confidence: 99%

“…Nevertheless, due to the lack of use of common standardised diagnostic tools between different TSE laboratories, it is today difficult to give a consistent description of the relative ratios of the multiple bands of PrP Sc from the atypical/Nor98 scrapie isolates. For example, the percentage of the total intensity represented by the lowest band that characterises the atypical/Nor98 scrapie cases varies considerably according to the mAb used and the proteinase K treatment of the isolate as illustrated by the differences observed in the literature [2,5,9,24]. It is quite plausible that most of the discrepancies could disappear by using common standardised diagnostic tools.…”

Section: Glycoprofile Of Prp Scmentioning

confidence: 99%

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Atypical/Nor98 scrapie: properties of the agent, genetics, and epidemiology

et al. 2008

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-Atypical/Nor98 scrapie cases in sheep were diagnosed for the first time in Norway in 1998. They are now identified in small ruminants in most European countries and represent an increasingly large proportion of the scrapie cases diagnosed in Europe. Atypical/Nor98 scrapie isolates have shown to be experimentally transmissible into transgenic mice and sheep but the properties of the TSE agent involved, like its biological and biochemical features, are so clearly distinct from the agent involved in classical scrapie that they have provided a challenging diagnostic for many years. No strain diversity has yet been identified among the atypical/Nor98 scrapie sample cases. The genetic predisposition of the sheep affected by atypical/Nor98 scrapie is almost inverted compared to classical scrapie, and the exact origin of this sporadic TSE strain is still speculative, but a spontaneous, non-contagious origin, like sporadic CreutzfeldtJakob disease in humans, can not be excluded. Further transmission and epidemiological studies are needed to better address this hypothesis. atypical scrapie / Nor98 / transmissible spongiform encephalopathies TSE / genetics / epidemiology

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Section: Identification Of New Prp Sc Fragmentsmentioning

confidence: 99%

Section: Deglycosylation Experimentsmentioning

confidence: 99%

Section: Prp Sc Cleavage Sitesmentioning

confidence: 99%

Section: Prp Sc Cleavage Sitesmentioning

confidence: 99%

Section: Glycoprofile Of Prp Scmentioning

confidence: 99%

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Atypical/Nor98 scrapie: properties of the agent, genetics, and epidemiology

et al. 2008

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A novel human disease with abnormal prion protein sensitive to protease

Gambetti

Dong

Yuan

et al. 2008

Annals of Neurology

253

280

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Objective: To report a novel prion disease characterized by distinct histopathological and immunostaining features, and associated with an abnormal isoform of the prion protein (PrP) that, contrary to the common prion diseases, is predominantly sensitive to protease digestion. Methods: Eleven subjects were investigated at the National Prion Disease Pathology Surveillance Center for clinical, histopathological, immunohistochemical, genotypical, and PrP characteristics. Results: Patients presented with behavioral and psychiatric manifestations on average at 62 years, whereas mean disease duration was 20 months. The type of spongiform degeneration, the PrP immunostaining pattern, and the presence of microplaques distinguished these cases from those with known prion diseases. Typical protease-resistant PrP was undetectable in the cerebral neocortex with standard diagnostic procedures. After enrichment, abnormal PrP was detected at concentrations 16 times lower than common prion diseases; it included nearly 4 times less protease-resistant PrP, which formed a distinct electrophoretic profile. The subjects examined comprised about 3% of sporadic cases evaluated by the National Prion Disease Pathology Surveillance Center. Although several subjects had family histories of dementia, no mutations were found in the PrP gene open reading frame. Interpretation: The distinct histopathological, PrP immunohistochemical, and physicochemical features, together with the homogeneous genotype, indicate that this is a previously unidentified type of disease involving the PrP, which we designated "protease-sensitive prionopathy" (or PSPr). Protease-sensitive prionopathy is not rare among prion diseases, and it may be even more prevalent than our data indicate because protease-sensitive prionopathy cases are likely also to be classified within the group of non-Alzheimer's dementias.

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High‐resolution differentiation of transmissible spongiform encephalopathy strains by quantitative N‐terminal amino acid profiling (N‐TAAP) of PK‐digested abnormal prion protein

et al. 2008

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New forms of transmissible spongiform encephalopathy (TSE) continue to be identified, and consequently sensitive differential diagnosis is increasingly important both for the management of disease in humans and livestock and in providing confidence in the safety of the food chain. TSE diseases are associated with accumulation of protease-resistant prion protein (PrP(Sc)) and detection of this marker protein is central to diagnosis. Proteolysis by proteinase K (PK) generates protease-resistant products (PrP(res)) with partially variable N-termini. The conformation(s) of PrP(Sc) and thus the points of PK cleavage are thought to be dependent on the strain of prion disease. Western blot (WB) analysis of PrP(res) gives characteristic migration patterns that can be used to diagnose TSEs, but the relatively low resolution of this technique limits its ability to differentiate certain disease strains. Mass spectrometry (MS) has the capability to resolve these various PK cleavage sites to the level of individual amino acid residues. In the present study multiple selected reaction monitoring (mSRM) was used to detect and quantify PrP(res) N-terminal tryptic peptides by MS and thus to define the N-terminal amino acid profiles (N-TAAPs) of PrP(res) characteristic for various TSEs in sheep. The fragmentation behaviour of the N-terminal tryptic peptides was studied to allow selection of the transitions specific for each peptide. Different PrP(res) preparation methods were evaluated and the most effective approach applied to differentiate the N-TAAPs corresponding to various sheep TSE isolates. Marked differences were identified between the N-TAAPs of bovine spongiform encephalopathy (BSE) and classical scrapie, and between classical scrapie and the experimental strains SSBP/1 and CH1641, thereby validating this approach as a means of TSE-strain specific diagnosis.

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Characterization of proteinase K-resistant N- and C-terminally truncated PrP in Nor98 atypical scrapie

Cited by 56 publications

References 45 publications

Atypical/Nor98 scrapie: properties of the agent, genetics, and epidemiology

Atypical/Nor98 scrapie: properties of the agent, genetics, and epidemiology

A novel human disease with abnormal prion protein sensitive to protease

High‐resolution differentiation of transmissible spongiform encephalopathy strains by quantitative N‐terminal amino acid profiling (N‐TAAP) of PK‐digested abnormal prion protein

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