scholarly journals Altered toxicity of the prion protein peptide PrP106–126 carrying the Ala117→Val mutation

2000 ◽  
Vol 346 (3) ◽  
pp. 785-791 ◽  
Author(s):  
David R. BROWN
Keyword(s):  
Point Mutation ◽  
Prion Protein ◽  
Prion Diseases ◽  
Point Mutations ◽  
Human Sequence ◽  
Gene Coding ◽  
Normal Human ◽  
Microtubule Formation ◽  
Β Sheet ◽  
Prion Protein Peptide

The inherited prion diseases such as Gerstmann-Sträussler-Scheinker syndrome (GSS) are linked to point mutations in the gene coding for the cellular isoform of the prion protein (PrPC). One particular point mutation A117V (Ala117 → Val) is linked to a variable pathology that usually includes deposition of neurofibrillary tangles. A prion protein peptide carrying this point mutation [PrP106-126(117V)] was generated and compared with a peptide based on the normal human sequence [PrP106-126(117A)]. The inclusion of this point mutation increased the toxicity of PrP106-126 which could be linked to an increased β-sheet content. An assay of microtubule formation in the presence of tau indicated that PrP106-126 decreased the rate of microtubule formation that could be related to the displacement of tau. PrP106-126 carrying the 117 mutation was more efficient at inhibiting microtubule formation. These results suggest a possible mechanism of toxicity for protein carrying this mutation via destabilization of the cytoskeleton and deposition of tau in filaments, as observed in GSS.

scholarly journals Assessment of the PrP c Amino-Terminal Domain in Prion Species Barriers

2016 ◽  
Vol 90 (23) ◽  
pp. 10752-10761 ◽  
Author(s):  
Kristen A. Davenport ◽  
Davin M. Henderson ◽  
Candace K. Mathiason ◽  
Edward A. Hoover
Keyword(s):  
Prion Protein ◽  
Bank Vole ◽  
Protein Misfolding ◽  
Prion Diseases ◽  
Full Length ◽  
Spongiform Encephalopathy ◽  
Amino Terminal ◽  
Terminal Domain ◽  
Human Sequence ◽  
Amino Terminal Domain

ABSTRACT Chronic wasting disease (CWD) in cervids and bovine spongiform encephalopathy (BSE) in cattle are prion diseases that are caused by the same protein-misfolding mechanism, but they appear to pose different risks to humans. We are interested in understanding the differences between the species barriers of CWD and BSE. We used real-time, quaking-induced conversion (RT-QuIC) to model the central molecular event in prion disease, the templated misfolding of the normal prion protein, PrP c , to a pathogenic, amyloid isoform, scrapie prion protein, PrP Sc . We examined the role of the PrP c amino-terminal domain (N-terminal domain [NTD], amino acids [aa] 23 to 90) in cross-species conversion by comparing the conversion efficiency of various prion seeds in either full-length (aa 23 to 231) or truncated (aa 90 to 231) PrP c . We demonstrate that the presence of white-tailed deer and bovine NTDs hindered seeded conversion of PrP c , but human and bank vole NTDs did the opposite. Additionally, full-length human and bank vole PrP c s were more likely to be converted to amyloid by CWD prions than were their truncated forms. A chimera with replacement of the human NTD by the bovine NTD resembled human PrP c . The requirement for an NTD, but not for the specific human sequence, suggests that the NTD interacts with other regions of the human PrP c to increase promiscuity. These data contribute to the evidence that, in addition to primary sequence, prion species barriers are controlled by interactions of the substrate NTD with the rest of the substrate PrP c molecule. IMPORTANCE We demonstrate that the amino-terminal domain of the normal prion protein, PrP c , hinders seeded conversion of bovine and white-tailed deer PrP c s to the prion forms, but it facilitates conversion of the human and bank vole PrP c s to the prion forms. Additionally, we demonstrate that the amino-terminal domain of human and bank vole PrP c s requires interaction with the rest of the molecule to facilitate conversion by CWD prions. These data suggest that interactions of the amino-terminal domain with the rest of the PrP c molecule play an important role in the susceptibility of humans to CWD prions.

scholarly journals Modulation of Prion Formation, Aggregation, and Toxicity by the Actin Cytoskeleton in Yeast

2006 ◽  
Vol 26 (2) ◽  
pp. 617-629 ◽  
Author(s):  
Elena E. Ganusova ◽  
Laura N. Ozolins ◽  
Srishti Bhagat ◽  
Gary P. Newnam ◽  
Renee D. Wegrzyn ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Prion Protein ◽  
De Novo ◽  
Prion Diseases ◽  
Actin Assembly ◽  
Yeast Prion ◽  
Cortical Actin ◽  
Gene Coding ◽  
Cortical Cytoskeleton ◽  
Yeast Deletion

ABSTRACT Self-perpetuating protein aggregates transmit prion diseases in mammals and heritable traits in yeast. De novo prion formation can be induced by transient overproduction of the corresponding prion-forming protein or its prion domain. Here, we demonstrate that the yeast prion protein Sup35 interacts with various proteins of the actin cortical cytoskeleton that are involved in endocytosis. Sup35-derived aggregates, generated in the process of prion induction, are associated with the components of the endocytic/vacuolar pathway. Mutational alterations of the cortical actin cytoskeleton decrease aggregation of overproduced Sup35 and de novo prion induction and increase prion-related toxicity in yeast. Deletion of the gene coding for the actin assembly protein Sla2 is lethal in cells containing the prion isoforms of both Sup35 and Rnq1 proteins simultaneously. Our data are consistent with a model in which cytoskeletal structures provide a scaffold for generation of large aggregates, resembling mammalian aggresomes. These aggregates promote prion formation. Moreover, it appears that the actin cytoskeleton also plays a certain role in counteracting the toxicity of the overproduced potentially aggregating proteins.

scholarly journals Expanding spectrum of prion diseases

2020 ◽  
Vol 4 (2) ◽  
pp. 155-167
Author(s):  
Jacob I. Ayers ◽  
Nick A. Paras ◽  
Stanley B. Prusiner
Keyword(s):  
Amino Acids ◽  
Nucleic Acid ◽  
Prion Protein ◽  
Prion Diseases ◽  
Lewy Body Dementia ◽  
Cellular Prion Protein ◽  
Chromosomal Gene ◽  
Scrapie Agent ◽  
Β Sheet ◽  
Translational Process

Prions were initially discovered in studies of scrapie, a transmissible neurodegenerative disease (ND) of sheep and goats thought to be caused by slow viruses. Once scrapie was transmitted to rodents, it was discovered that the scrapie pathogen resisted inactivation by procedures that modify nucleic acids. Eventually, this novel pathogen proved to be a protein of 209 amino acids, which is encoded by a chromosomal gene. After the absence of a nucleic acid within the scrapie agent was established, the mechanism of infectivity posed a conundrum and eliminated a hypothetical virus. Subsequently, the infectious scrapie prion protein (PrPSc) enriched for β-sheet was found to be generated from the cellular prion protein (PrPC) that is predominantly α-helical. The post-translational process that features in nascent prion formation involves a templated conformational change in PrPC that results in an infectious copy of PrPSc. Thus, prions are proteins that adopt alternative conformations, which are self-propagating and found in organisms ranging from yeast to humans. Prions have been found in both Alzheimer's (AD) and Parkinson's (PD) diseases. Mutations in APP and α-synuclein genes have been shown to cause familial AD and PD. Recently, AD was found to be a double prion disorder: both Aβ and tau prions feature in this ND. Increasing evidence argues for α-synuclein prions as the cause of PD, multiple system atrophy, and Lewy body dementia.

Biophysical characterization of oligomerization and fibrillization of the G131V pathogenic mutant of human prion protein

2019 ◽  
Author(s):  
Meilan Zhang ◽  
Haoran Zhang ◽  
Hongwei Yao ◽  
Chenyun Guo ◽  
Donghai Lin
Keyword(s):  
Prion Protein ◽  
Tertiary Structure ◽  
Prion Proteins ◽  
Prion Diseases ◽  
Guanidine Hydrochloride ◽  
Point Mutations ◽  
Cellular Prion Protein ◽  
Human Prion Protein ◽  
Α Helix ◽  
Conformational Conversion

Abstract The pathogenesis of fatal neurodegenerative prion diseases is closely associated with the conversion of α-helix-rich cellular prion protein into β-sheet-rich scrapie form. Pathogenic point mutations of prion proteins usually promote the conformational conversion and trigger inherited prion diseases. The G131V mutation of human prion protein (HuPrP) was identified to be involved in Gerstmann–Sträussler–Scheinker syndrome. Few studies have been carried out to address the pathogenesis of the G131V mutant. Here, we addressed the effects of the G131V mutation on oligomerization and fibrillization of the full-length HuPrP(23–231) and truncated HuPrP(91–231) proteins. The G131V mutation promotes the oligomerization but alleviates the fibrillization of HuPrP, implying that the oligomerization might play a crucial role in the pathogenic mechanisms of the G131V mutant. Moreover, the flexible N-terminal fragment in either the wild-type or the G131V mutant HuPrP increases the oligomerization tendencies but decreases the fibrillization tendencies. Furthermore, this mutation significantly alters the tertiary structure of human PrPC and might distinctly change the conformational conversion tendency. Interestingly, both guanidine hydrochloride denaturation and thermal denaturation experiments showed that the G131V mutation does not significantly change the thermodynamic stabilities of the HuPrP proteins. This work may be of benefit to a mechanistic understanding of the conformational conversion of prion proteins and also provide clues for the prevention and treatment of prion diseases.

scholarly journals Heterogeneity and Architecture of Pathological Prion Protein Assemblies: Time to Revisit the Molecular Basis of the Prion Replication Process?

Viruses ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 429 ◽  
Author(s):  
Angélique Igel-Egalon ◽  
Jan Bohl ◽  
Mohammed Moudjou ◽  
Laetitia Herzog ◽  
Fabienne Reine ◽  
...  
Keyword(s):  
Prion Protein ◽  
Current Knowledge ◽  
Prion Diseases ◽  
Structural Heterogeneity ◽  
Cellular Prion Protein ◽  
Protein Assemblies ◽  
Replication Process ◽  
Data Points ◽  
Autocatalytic Process ◽  
Β Sheet

Prions are proteinaceous infectious agents responsible for a range of neurodegenerative diseases in animals and humans. Prion particles are assemblies formed from a misfolded, β-sheet rich, aggregation-prone isoform (PrPSc) of the host-encoded cellular prion protein (PrPC). Prions replicate by recruiting and converting PrPC into PrPSc, by an autocatalytic process. PrPSc is a pleiomorphic protein as different conformations can dictate different disease phenotypes in the same host species. This is the basis of the strain phenomenon in prion diseases. Recent experimental evidence suggests further structural heterogeneity in PrPSc assemblies within specific prion populations and strains. Still, this diversity is rather seen as a size continuum of assemblies with the same core structure, while analysis of the available experimental data points to the existence of structurally distinct arrangements. The atomic structure of PrPSc has not been elucidated so far, making the prion replication process difficult to understand. All currently available models suggest that PrPSc assemblies exhibit a PrPSc subunit as core constituent, which was recently identified. This review summarizes our current knowledge on prion assembly heterogeneity down to the subunit level and will discuss its importance with regard to the current molecular principles of the prion replication process.

scholarly journals Comparative analysis of heparin affecting the biochemical properties of chicken and murine prion proteins

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247248
Author(s):  
Li-Juan Wang ◽  
Xiao-Dan Gu ◽  
Xiao-Xiao Li ◽  
Liang Shen ◽  
Hong-Fang Ji
Keyword(s):  
Prion Protein ◽  
Conformational Changes ◽  
Protein Misfolding ◽  
Prion Diseases ◽  
Mammalian Species ◽  
Biochemical Properties ◽  
Cellular Prion Protein ◽  
Great Loss ◽  
Heparin Concentration ◽  
Β Sheet

The conversion of cellular prion protein (PrPC) to disease-provoking conformer (PrPSc) is crucial in the pathogenesis of prion diseases. Heparin has been shown to enhance mammalian prion protein misfolding. As spontaneous prion disease has not been reported in non-mammalian species, such as chicken, it is interesting to explore the influence of heparin on the conversion of chicken prion protein (ChPrP). Herein, we investigated the influences of heparin on biochemical properties of full-length recombinant ChPrP, with murine prion protein (MoPrP) as control. The results showed that at low heparin concentration (10 μg/mL), a great loss of solubility was observed for both MoPrP and ChPrP using solubility assays. In contrast, when the concentration of heparin was high (30 μg/mL), the solubility of MoPrP and ChPrP both decreased slightly. Using circular dichroism, PK digestion and transmission electron microscopy, significantly increased β-sheet content, PK resistance and size of aggregates were observed for MoPrP interacted with 30 μg/mL heparin, whereas 30 μg/mL heparin-treated ChPrP showed less PK resistance and slight increase of β-sheet structure. Therefore, heparin can induce conformational changes in both MoPrP and ChPrP and the biochemical properties of the aggregates induced by heparin could be modified by heparin concentration. These results highlight the importance of concentration of cofactors affecting PrP misfolding.

scholarly journals NMR characterization of the pH 4 β-intermediate of the prion protein: the N-terminal half of the protein remains unstructured and retains a high degree of flexibility

2006 ◽  
Vol 401 (2) ◽  
pp. 533-540 ◽  
Author(s):  
Denis B. D. O'Sullivan ◽  
Christopher E. Jones ◽  
Salama R. Abdelraheim ◽  
Andrew R. Thompsett ◽  
Marcus W. Brazier ◽  
...  
Keyword(s):  
Prion Protein ◽  
Prion Diseases ◽  
Molten Globule ◽  
Three Dimensional ◽  
Molecular Size ◽  
Native Form ◽  
C Terminus ◽  
Nmr Characterization ◽  
Helical Protein ◽  
Β Sheet

Prion diseases are associated with the misfolding of the PrP (prion protein) from a largely α-helical isoform to a β-sheet-rich oligomer. CD has shown that lowering the pH to 4 under mildly denaturing conditions causes recombinant PrP to convert from an α-helical protein into one that contains a high proportion of β-sheet-like conformation. In the present study, we characterize this soluble pH 4 folding intermediate using NMR. 15N-HSQC (heteronuclear single-quantum correlation) studies with mPrP (mouse PrP)-(23–231) show that a total of 150 dispersed amide signals are resolved in the native form, whereas only 65 amide signals with little chemical shift dispersion are observable in the pH 4 form. Three-dimensional 15N-HSQC-TOCSY and NOESY spectra indicate that the observable residues are all assigned to amino acids in the N-terminus: residues 23–118. 15N transverse relaxation measurements indicate that these N-terminal residues are highly flexible with additional fast motions. These observations are confirmed via the use of truncated mPrP-(112–231), which shows only 16 15N-HSQC amide peaks at pH 4. The loss of signals from the C-terminus can be attributed to line broadening due to an increase in the molecular size of the oligomer or exchange broadening in a molten-globule state.

scholarly journals Amyloid fibrils from the N-terminal prion protein fragment are infectious

2016 ◽  
Vol 113 (48) ◽  
pp. 13851-13856 ◽  
Author(s):  
Jin-Kyu Choi ◽  
Ignazio Cali ◽  
Krystyna Surewicz ◽  
Qingzhong Kong ◽  
Pierluigi Gambetti ◽  
...  
Keyword(s):  
Prion Protein ◽  
Prion Disease ◽  
Amyloid Fibrils ◽  
Prion Diseases ◽  
Strong Support ◽  
Proteinase K ◽  
Molecular Architecture ◽  
Core Mapping ◽  
Scrapie Strains ◽  
Β Sheet

Recombinant C-terminally truncated prion protein PrP23-144 (which corresponds to the Y145Stop PrP variant associated with a Gerstmann–Sträussler–Scheinker-like prion disease) spontaneously forms amyloid fibrils with a parallel in-register β-sheet architecture and β-sheet core mapping to residues ∼112–139. Here we report that mice (bothtga20and wild type) inoculated with a murine (moPrP23-144) version of these fibrils develop clinical prion disease with a 100% attack rate. Remarkably, even though fibrils in the inoculum lack the entire C-terminal domain of PrP, brains of clinically sick mice accumulate longer proteinase K-resistant (PrPres) fragments of ∼17–32 kDa, similar to those observed in classical scrapie strains. Shorter, Gerstmann–Sträussler–Scheinker-like PrPresfragments are also present. The evidence that moPrP23-144 amyloid fibrils generated in the absence of any cofactors are bona fide prions provides a strong support for the protein-only hypothesis of prion diseases in its pure form, arguing against the notion that nonproteinaceous cofactors are obligatory structural components of all infectious prions. Furthermore, our finding that a relatively short β-sheet core of PrP23-144 fibrils (residues ∼112–139) with a parallel in-register organization of β-strands is capable of seeding the conversion of full-length prion protein to the infectious form has important implications for the ongoing debate regarding structural aspects of prion protein conversion and molecular architecture of mammalian prions.

Assembly of natural and recombinant prion protein into fibrils

2005 ◽  
Vol 386 (6) ◽  
pp. 569-580 ◽  
Author(s):  
Karl-Werner Leffers ◽  
Holger Wille ◽  
Jan Stöhr ◽  
Erika Junger ◽  
Stanley B. Prusiner ◽  
...  
Keyword(s):  
Prion Protein ◽  
Amyloid Fibrils ◽  
Prion Diseases ◽  
Sodium Dodecyl ◽  
Limited Proteolysis ◽  
Fibril Formation ◽  
Full Length ◽  
Low Concentrations ◽  
Recombinant Prion Protein ◽  
Β Sheet

AbstractThe conversion of the α-helical, cellular isoform of the prion protein (PrPC) to the insoluble, β-sheet-rich, infectious, disease-causing isoform (PrPSc) is the fundamental event in the prion diseases. The C-terminal fragment of PrPSc(PrP 27–30) is formed by limited proteolysis and retains infectivity. Unlike full-length PrPSc, PrP 27–30 polymerizes into rod-shaped structures with the ultra-structural and tinctorial properties of amyloid. To study the folding of PrP, both with respect to the formation of PrPScfrom PrPCand the assembly of rods from PrP 27–30, we solubilized Syrian hamster (sol SHa) PrP 27–30 in low concentrations (0.2%) of sodium dodecyl sulfate (SDS) under conditions previously used to study the structural transitions of this protein. Sol SHaPrP 27–30 adopted a β-sheet-rich structure at SDS concentrations between 0.02% and 0.04% and remained soluble. Here we report that NaCl stabilizes SHaPrP 27–30 in a soluble, β-sheet-rich state that allows fibril assembly to proceed over several weeks. Under these conditions, fibril formation occurred not only with sol PrP 27–30, but also with native SHaPrPC. Addition of sphingolipids seems to increase fibril growth. When recombinant (rec) SHaPrP(90–231) was exposed to low concentrations of SDS, similar to those used to polymerize sol SHaPrP 27–30 in the presence of 250 mM NaCl, fibril formation occurred regularly. When fibrils formed from PrP 27–30 or PrPCwere bioassayed in transgenic mice overexpressing full-length SHaPrP, no infectivity was obtained, whereas amyloid fibrils formed of rec mouse PrP(89–230) were infectious. At present, it cannot be determined whether the lack of infectivity is caused by a difference in the structure of the fibrils or in the bioassay conditions.

An Engineered PrPsc-like Molecule from the Chimera of Mammalian Prion Protein and Yeast Ure2p Prion-inducing Domain

2004 ◽  
Vol 36 (2) ◽  
pp. 128-132
Author(s):  
Shao-Man Yin ◽  
Man-Sun Sy ◽  
Po Tien
Keyword(s):  
Prion Protein ◽  
Amyloid Fibrils ◽  
Physiological Condition ◽  
Prion Diseases ◽  
Cd Spectroscopy ◽  
Conversion Process ◽  
Fibrillar Morphology ◽  
Thioflavine T ◽  
Conformational Conversion ◽  
Β Sheet

Abstract Production of the pathogenic prion isoform PrPsc-like molecules is thought to be useful for understanding the mysterious mechanism of conformational conversion process of prion diseases and proving the “protein-only” hypothesis. In this report, an engineered PrPsc-like conformation was produced from a chimera of mammalian bovine prion protein (bPrP) and yeast Ure2p prion-inducing domain (UPrD). Compared with the normal form of bPrP, the engineered recombinant protein, termed bPrP-UPrD, spontaneously aggregated into ordered fibrils under physiological condition, displaying amyloid-like characteristics, such as fibrillar morphology, birefringence upon binding to Congo red and increased fluorescence intensity with Thioflavine T. Limited resistance to protease K digestion and CD spectroscopy experiments suggested that the structure of bPrP-UPrD had been changed, and adopted a new, high content β-sheet conformation during the fibrils formation. Moreover, bPrP-UPrD amyloid fibrils could recruit more soluble forms into the aggregates. Therefore, the engineered molecules could mimic significant behaviors of PrPsc and will be helpful for further understanding the mechanism of conformational conversion process.

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