Proteinase K-sensitive disease-associated ovine prion protein revealed by conformation-dependent immunoassay

Thackray, Alana M.; Hopkins, Lee; Mathiason, Candace K.

doi:10.1042/bj20061264

Cited by 72 publications

(71 citation statements)

References 55 publications

(84 reference statements)

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“…Nevertheless, the amount of neuronal loss and astrocytosis was similar in both groups in the cortex, hippocampus, cerebellum, and brainstem, indicating that residual PrP res remains capable of causing neurodegeneration and death. The existence of proteinase-sensitive infectious prion protein (PrP Sc ) has been reported by others, and copper is known to influence the proteinase resistance of PrP (41)(42)(43). It is possible that, although accumulated PrP Sc in a copper-deficient environment may be more sensitive to digestion by proteinases, it may remain toxic enough to cause disease.…”

Section: Discussionmentioning

confidence: 99%

Disruption of copper homeostasis due to a mutation of Atp7a delays the onset of prion disease

Siggs

Cruite

et al. 2012

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

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Copper influences the pathogenesis of prion disease, but whether it is beneficial or detrimental remains controversial. Copper homeostasis is also essential for normal physiology, as highlighted by the spectrum of diseases caused by disruption of the copper transporting enzymes ATP7A and ATP7B. Here, by using a forward genetics approach in mice, we describe the isolation of three alleles of Atp7a , each with different phenotypic consequences. The mildest of the three, Atp7a brown , was insufficient to cause lethality in hemizygotes or mottling of the coat in heterozygotes, but did lead to coat hypopigmentation and reduced copper content in the brains of hemizygous males. When challenged with Rocky Mountain Laboratory scrapie, the onset of prion disease was delayed in Atp7a brown mice, and significantly less proteinase-resistant prion protein was found in the brains of moribund Atp7a brown mice compared with WT littermates. Our results establish that ATP7A-mediated copper homeostasis is important for the formation of pathogenic proteinase-resistant prion protein.

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Section: Discussionmentioning

confidence: 99%

Disruption of copper homeostasis due to a mutation of Atp7a delays the onset of prion disease

Siggs

Cruite

et al. 2012

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

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“…Protease-sensitive PrP species are still poorly defined and range from PK-sensitive patterns resembling PrP Sc digestion patterns and different sized multimers to completely novel digestion patterns including small fragments (Cronier et al, 2008;Gambetti et al, 2008;Safar et al, 1998;Sajnani et al, 2012;Thackray et al, 2007;Tzaban et al, 2002). However, here, protease sensitivity was defined as a PKdegradable PrP protein signal that was detectable using defined anti-PrP antibodies after Western blotting.…”

Section: Discussionmentioning

confidence: 99%

Protease-sensitive prion species in neoplastic spleens of prion-infected mice with uncoupling of PrPSc and prion infectivity

Krasemann

Neumann

Szalay

et al. 2013

Journal of General Virology

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Prion diseases are fatal neurodegenerative disorders. An important step in disease pathophysiology is the conversion of cellular prion protein (PrPC) to disease-associated misfolded conformers (PrPSc). These misfolded PrP variants are a common component of prion infectivity and are detectable in diseased brain and lymphoreticular organs such as spleen. In the latter, PrPSc is thought to replicate mainly in follicular dendritic cells within spleen follicles. Although the presence of PrPSc is a hallmark for prion disease and serves as a main diagnostic criterion, in certain instances the amount of PrPSc does not correlate well with neurotoxicity or prion infectivity. Therefore, it has been proposed that prions might be a mixture of different conformers and aggregates with differing properties. This study investigated the impact of disruption of spleen architecture by neoplasia on the abundance of different PrP species in spleens of prion-infected mice. Although follicular integrity was completely disturbed, titres of prion infectivity in neoplastic spleens were not significantly altered, yet no protease-resistant PrPSc was detectable. Instead, unique protease-sensitive prion species could be detected in neoplastic spleens. These results indicate the dissociation of PrPSc and prion infectivity and showed the presence of non-PrPSc PrP species in spleen with divergent biochemical properties that become apparent after tissue architecture disruption.

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“…CDI has been used to show that a significant proportion of PrP Sc in the brains of scrapie-infected hamsters is actually susceptible to proteolytic degradation (Safar et al, 1998). In a subsequent study, up to 90 % of the PrP Sc present in the brains of patients affected with sCJD was reported to be PK-sensitive (Thackray et al, 2007). Moreover, various prion strains propagated in hamsters were reported to be distinguishable based on the ratio of PK-resistant PrP Sc to PK-sensitive PrP Sc (Safar et al, 1998).…”

Section: An Alternative Approach That Also Distinguishes Prpmentioning

confidence: 99%

Comparison of the level, distribution and form of disease-associated prion protein in variant and sporadic Creutzfeldt-Jakob diseased brain using conformation-dependent immunoassay and Western blot

Choi

Gröner

Ironside

et al. 2010

Journal of General Virology

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Disease-associated prion protein (PrP Sc ) can be distinguished from the cellular isoform (PrP C ) by conformation-dependent immunoassay (CDI). This technique exploits the presence of an epitope, accessible in PrP C , but only unmasked by denaturation in PrP Sc . In this study, we investigated PrP Sc in different brain regions in variant and sporadic Creutzfeldt-Jakob disease (CJD) by using CDI, and directly compared the results with those obtained using the more commonly employed protease digestion and Western blotting. In general, there was good agreement between the results, although there were certain discrepancies in relative abundance when the regional distribution in variant CJD cases was considered. The results largely confirmed the previously described targeting of different brain regions by variant and sporadic CJD. Additionally, the combination of protease digestion and CDI detection demonstrated, for the first time, the presence of PrP Sc in variant CJD brains that is susceptible to proteolysis under standard conditions.Prion diseases are characterized by the conversion of the host-encoded prion protein (PrP C ) into a disease-associated isoform (PrP Sc ), which is thought to be the main component of the infectious agent or prion (Bolton et al., 1982;Prusiner, 1982Prusiner, , 1998. The conversion of a-helical PrP C into b-sheet-rich PrP Sc accompanies change in biochemical properties, such as protease resistance and detergent solubility (Meyer et al., 1986;Caughey et al., 1991;Pan et al., 1993). The proteolytic treatment of PrP Sc leaves an N-terminally truncated, proteinase K (PK)-resistant core fragment of PrP Sc (termed PrP res ) (Oesch et al., 1985). In human prion diseases, the presence of PrP Sc in a sample of affected tissue (usually grey matter enriched cerebral cortex or cerebellum) has been most commonly demonstrated by detecting PrP res by Western blotting. Variant Creutzfeldt-Jakob disease (vCJD) can be distinguished from sporadic CJD (sCJD) based on the PrP res type as determined by the size of the N-terminally truncated non-glycosylated fragment and the proportion of three possible glycoforms (Collinge et al., 1996;Head et al., 2004). Additionally, the type of PrP res in combination with genotype at PRNP codon 129 has been proposed as the basis of a molecular classification system accounting for the marked clinico-pathological variability of sCJD (Parchi et al., 1999). An alternative approach that also distinguishes PrPSc from PrP C is conformation-dependent immunoassay (CDI), which is based on a conformational transition in the N terminus of PrP during the formation of PrP Sc (Peretz et al., 1997). CDI was initially developed in the form of a direct ELISA using the mAb 3F4 that binds to aa 109-112 of human PrP (Safar et al., 1998). The epitope of mAb 3F4 was found not to be accessible in native PrP Sc , in contrast to its accessibility in PrP C , and was shown to become exposed in PrP Sc after denaturation (Safar et al., 1998). The introduction of sodium phosphotungstic acid (NaPTA) p...

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Proteinase K-sensitive disease-associated ovine prion protein revealed by conformation-dependent immunoassay

Cited by 72 publications

References 55 publications

Disruption of copper homeostasis due to a mutation of Atp7a delays the onset of prion disease

Disruption of copper homeostasis due to a mutation of Atp7a delays the onset of prion disease

Protease-sensitive prion species in neoplastic spleens of prion-infected mice with uncoupling of PrPSc and prion infectivity

Comparison of the level, distribution and form of disease-associated prion protein in variant and sporadic Creutzfeldt-Jakob diseased brain using conformation-dependent immunoassay and Western blot

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