Identification of Two Prion Protein Regions That Modify Scrapie Incubation Time

Supattapone, Surachai; Muramoto, Tamaki; Legname, Giuseppe; Mehlhorn, Ingrid; Cohen, Fred E.; DeArmond, Stephen J.; Prusiner, Stanley B.; Scott, Michael

doi:10.1128/jvi.75.3.1408-1413.2001

Cited by 102 publications

(115 citation statements)

References 35 publications

(31 reference statements)

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“…2). Using the PTA precipitation method, we partially purified 301V prions from FVB mice and MoSP1 prions from transgenic (Tg) mice overexpressing MoPrP(89-231), designated Tg9949 mice (18). The 301V strain was derived from bovine spongiform encephalopathy prions passaged through mice (19).…”

Section: Prion-infected Brain Homogenates Can Seed Amyloid Formationmentioning

confidence: 99%

Prion detection by an amyloid seeding assay

Colby

Zhang²,

Wang³

et al. 2007

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

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Polymerization of recombinant prion protein (recPrP), which was produced in bacteria, into amyloid fibers was accompanied by the acquisition of prion infectivity. We report here that partially purified preparations of prions seed the polymerization of recPrP into amyloid as detected by a fluorescence shift in the dye Thioflavin T. Our amyloid seeding assay (ASA) detected PrP Sc , the sole component of the prion, in brain samples from humans with sporadic Creutzfeldt–Jakob disease, as well as in rodents with experimental prion disease. The ASA detected a variety of prion strains passaged in both mice and hamsters. The sensitivity of the ASA varied with strain type; for hamster Sc237 prions, the limit of detection was ≈1 fg. Some prion strains consist largely of protease-sensitive PrP Sc (sPrP Sc ), and these strains were readily detected by ASA. Our studies show that the ASA provides an alternative methodology for detecting both sPrP Sc and protease-resistant PrP Sc that does not rely on protease digestion or immunodetection.

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Section: Prion-infected Brain Homogenates Can Seed Amyloid Formationmentioning

confidence: 99%

Prion detection by an amyloid seeding assay

Colby

Zhang²,

Wang³

et al. 2007

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

Self Cite

207

185

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“…1 A). At the N terminus, we found that HuM-E123, which recognizes the region of MoPrP (30)(31)(32)(33)(34)(35)(36)(37), bound weakly, and that HuM-E149, a HuM-Fab specific to the octarepeat region, bound even more weakly than HuM-E123 (Fig. 1 A).…”

Section: Immunochemical Characterization Of Wtmoprp-fc By Using Hum-fmentioning

confidence: 94%

Prion and doppel proteins bind to granule cells of the cerebellum

Legname

Nelken²,

Guan³

et al. 2002

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

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“…The isolation of 301V (44), Me7 (45), RML (46), and synthetic prions (18) has been described. Tg9949 and Tg4053 mice have been described (47,48). CD-1 and FVB mice were purchased from Charles River Breeding Laboratories (Wilmington, MA).…”

Section: Methodsmentioning

confidence: 99%

Continuum of prion protein structures enciphers a multitude of prion isolate-specified phenotypes

Legname

Nguyen

Peretz

et al. 2006

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

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On passaging synthetic prions, two isolates emerged with incubation times differing by nearly 100 days. Using conformational-stability assays, we determined the guanidine hydrochloride (Gdn⅐HCl) concentration required to denature 50% of disease-causing prion protein (PrP Sc ) molecules, denoted as the [Gdn⅐HCl]1/2 value. For the two prion isolates enciphering shorter and longer incubation times, [Gdn⅐HCl]1/2 values of 2.9 and 3.7 M, respectively, were found. Intrigued by this result, we measured the conformational stabilities of 30 prion isolates from synthetic and naturally occurring sources that had been passaged in mice. When the incubation times were plotted as a function of the [Gdn⅐HCl] 1/2 values, a linear relationship was found with a correlation coefficient of 0.93. These findings demonstrate that (i) less stable prions replicate more rapidly than do stable prions, and (ii) a continuum of PrP Sc structural states enciphers a multitude of incubation-time phenotypes. Our data argue that cellular machinery must exist for propagating a large number of different PrP Sc conformers, each of which enciphers a distinct biological phenotype as reflected by a specific incubation time. The biophysical explanation for the unprecedented plasticity of PrP Sc remains to be determined.conformational stability ͉ memory ͉ strain ͉ synthetic prions ͉ tertiary structure

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Identification of Two Prion Protein Regions That Modify Scrapie Incubation Time

Cited by 102 publications

References 35 publications

Prion detection by an amyloid seeding assay

Prion detection by an amyloid seeding assay

Prion and doppel proteins bind to granule cells of the cerebellum

Continuum of prion protein structures enciphers a multitude of prion isolate-specified phenotypes

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