Glycan-Controlled Epitopes of Prion Protein Include a Major Determinant of Susceptibility to Sheep Scrapie

Moudjou, Mohammed; Tréguer, Eric; Rezaei, Human; Sabuncu, Elifsu; Neuendorf, Erdmute; Groschup, Martin H.; Grosclaude, Jeanne; Laude, Hubert

doi:10.1128/jvi.78.20.11449.2004

Cited by 6 publications

(7 citation statements)

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“…Blots were immediately incubated with PrP antibody Sha31 (Spi‐Bio Inc; 1/30,000 in 0.5% TBST) or were blocked with 5% skim milk in 0.1% TBST for 1 h at room temperature and incubated with primary antibody at 4°C overnight (PrP antibodies: 9A2 1/4,000, 12B2 1/8,500, SAF83 (Cayman), VRQ61 1/5,000, and 2D6 1/10,000, 1A6 1/5,000). 9A2 and 12B2 were gifts from Dr. Jan Langeveld (Langeveld et al , ), VRQ61 and 2D6 from Dr. Human Rezaei (Moudjou et al , ), and 1A6 was produced in‐house. Membranes were subsequently incubated with secondary antibody at 1/10,000 (Bio‐Rad) for 2 h at room temperature and visualized using ECL (Pierce).…”

Section: Methodsmentioning

confidence: 99%

Octarepeat region flexibility impacts prion function, endoproteolysis and disease manifestation

et al. 2015

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The cellular prion protein (PrPC) comprises a natively unstructured N-terminal domain, including a metal-binding octarepeat region (OR) and a linker, followed by a C-terminal domain that misfolds to form PrPSc in Creutzfeldt-Jakob disease. PrPC β-endoproteolysis to the C2 fragment allows PrPSc formation, while α-endoproteolysis blocks production. To examine the OR, we used structure-directed design to make novel alleles, ‘S1’ and ‘S3’, locking this region in extended or compact conformations, respectively. S1 and S3 PrP resembled WT PrP in supporting peripheral nerve myelination. Prion-infected S1 and S3 transgenic mice both accumulated similar low levels of PrPSc and infectious prion particles, but differed in their clinical presentation. Unexpectedly, S3 PrP overproduced C2 fragment in the brain by a mechanism distinct from metal-catalysed hydrolysis reported previously. OR flexibility is concluded to impact diverse biological endpoints; it is a salient variable in infectious disease paradigms and modulates how the levels of PrPSc and infectivity can either uncouple or engage to drive the onset of clinical disease.

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Section: Methodsmentioning

confidence: 99%

Octarepeat region flexibility impacts prion function, endoproteolysis and disease manifestation

et al. 2015

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“…On the blot probed with the V14 antibody that specifically detects mono181 [11,12], the PrP band migrating at ,24-26 kDa corresponding to mono181 was detected in sCJD in addition to the band migrating at ,19-20 kDa corresponding to the unglycosylated PrP (Fig. 2B).…”

Section: Both Inherited Cjd V180i and Sporadic Vpspr Exhibit No Diglycosylated Prp Resmentioning

confidence: 99%

“…To investigate whether and how the two individual N181 and N197 sites are associated with the lack of the diglycosylated PrP res in fCJD V180I and VPSPr, we probed PrP treated with PK or PK plus PNGase F using V14 and Bar209 antibodies that have been demonstrated to distinguish mono181 and mono197 but not diand unglycosylated PrP (Fig. S1 and S2) [11][12][13][14]. On the blot probed with the Bar209 antibody that specifically detects mono197, both mono197 and unglycosylated PrP migrating at ,26-28 kDa and ,19-20 kDa, respectively, were detected in all samples treated with PK alone except for fCJD T183A that had a significantly under-represented unglycosylated PrP (Fig.…”

Section: Both Inherited Cjd V180i and Sporadic Vpspr Exhibit No Diglycosylated Prp Resmentioning

confidence: 99%

Correction: Glycoform-Selective Prion Formation in Sporadic and Familial Forms of Prion Disease

Xiao¹,

Yuan²,

Haı̈k³

et al. 2013

PLoS ONE

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The four glycoforms of the cellular prion protein (PrP C ) variably glycosylated at the two N-linked glycosylation sites are converted into their pathological forms (PrP Sc ) in most cases of sporadic prion diseases. However, a prominent molecular characteristic of PrP Sc in the recently identified variably protease-sensitive prionopathy (VPSPr) is the absence of a diglycosylated form, also notable in familial Creutzfeldt-Jakob disease (fCJD), which is linked to mutations in PrP either from Val to Ile at residue 180 (fCJD V180I ) or from Thr to Ala at residue 183 (fCJD T183A ). Here we report that fCJD V180I , but not fCJD T183A , exhibits a proteinase K (PK)-resistant PrP (PrP res ) that is markedly similar to that observed in VPSPr, which exhibits a five-step ladder-like electrophoretic profile, a molecular hallmark of VPSPr. Remarkably, the absence of the diglycosylated PrP res species in both fCJD V180I and VPSPr is likewise attributable to the absence of PrP res glycosylated at the first N-linked glycosylation site at residue 181, as in fCJD T183A . In contrast to fCJD T183A , both VPSPr and fCJD V180I exhibit glycosylation at residue 181 on di-and monoglycosylated (mono181) PrP prior to PK-treatment. Furthermore, PrP V180I with a typical glycoform profile from cultured cells generates detectable PrP res that also contains the diglycosylated PrP in addition to mono-and unglycosylated forms upon PK-treatment. Taken together, our current in vivo and in vitro studies indicate that sporadic VPSPr and familial CJD V180I share a unique glycoform-selective prion formation pathway in which the conversion of diglycosylated and mono181 PrP C to PrP Sc is inhibited, probably by a dominant-negative effect, or by other co-factors.

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“…Several factors, alone or in combination, might account for this genetic modulation. Variations in conformations and glycosylation patterns between allelic variants have been suggested to modulate scrapie susceptibility by altering solvent accessibility (19,20). PrP C variants susceptible to classical scrapie have been shown to have longer cellular half-lives (21) and higher plasma concentrations (22), when compared to resistant variants, which might give them more opportunities to convert and oligomerize.…”

Section: Introductionmentioning

confidence: 99%

Microsecond Unfolding Kinetics of Sheep Prion Protein Reveals an Intermediate that Correlates with Susceptibility to Classical Scrapie

Chen

Wedemeyer

et al. 2011

Biophysical Journal

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The microsecond folding and unfolding kinetics of ovine prion proteins (ovPrP) were measured under various solution conditions. A fragment comprising residues 94-233 of the full-length ovPrP was studied for four variants with differing susceptibilities to classical scrapie in sheep. The observed biexponential unfolding kinetics of ovPrP provides evidence for an intermediate species. However, in contrast to previous results for human PrP, there is no evidence for an intermediate under refolding conditions. Global analysis of the kinetic data, based on a sequential three-state mechanism, quantitatively accounts for all folding and unfolding data as a function of denaturant concentration. The simulations predict that an intermediate accumulates under both folding and unfolding conditions, but is observable only in unfolding experiments because the intermediate is optically indistinguishable from the native state. The relative population of intermediates in two ovPrP variants, both transiently and under destabilizing equilibrium conditions, correlates with their propensities for classical scrapie. The variant susceptible to classical scrapie has a larger population of the intermediate state than the resistant variant. Thus, the susceptible variant should be favored to undergo the PrP(C) to PrP(Sc) conversion and oligomerization.

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Glycan-Controlled Epitopes of Prion Protein Include a Major Determinant of Susceptibility to Sheep Scrapie

Cited by 6 publications

References 0 publications

Octarepeat region flexibility impacts prion function, endoproteolysis and disease manifestation

Octarepeat region flexibility impacts prion function, endoproteolysis and disease manifestation

Correction: Glycoform-Selective Prion Formation in Sporadic and Familial Forms of Prion Disease

Microsecond Unfolding Kinetics of Sheep Prion Protein Reveals an Intermediate that Correlates with Susceptibility to Classical Scrapie

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