Truncated Forms of the Human Prion Protein in Normal Brain and in Prion Diseases

AbstractTransmissible spongiform encephalopathies (TSEs) are fatal neurodegenerative disorders associated with the misfolding and aggregation of the human prion protein (huPrP). Despite efforts into investigating the process of huPrP aggregation, the mechanisms triggering its misfolding remain elusive. A number of TSE-associated mutations of huPrP have been identified, but their role at the onset and progression of prion diseases is unclear. Here we report the NMR assignments of the C-terminal globular domain of the wild type huPrP and the pathological mutant T183A. The differences in chemical shifts between the two variants reveal conformational alterations in some structural elements of the mutant, whereas the analyses of secondary shifts and random coil index provide indications on the putative mechanisms of misfolding of T183A huPrP.

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Structural effects of the highly protective V127 polymorphism on human prion protein

Communications Biology ◽

10.1038/s42003-020-01126-6 ◽

2020 ◽

Vol 3 (1) ◽

Author(s):

Laszlo L. P. Hosszu ◽

Rebecca Conners ◽

Daljit Sangar ◽

Mark Batchelor ◽

Elizabeth B. Sawyer ◽

...

Keyword(s):

Prion Protein ◽

Prion Disease ◽

Structural Changes ◽

Prion Diseases ◽

Single Point ◽

Structural Basis ◽

Single Point Mutation ◽

Solution Dynamics ◽

Human Prion Protein ◽

Prion Transmission

AbstractPrion diseases, a group of incurable, lethal neurodegenerative disorders of mammals including humans, are caused by prions, assemblies of misfolded host prion protein (PrP). A single point mutation (G127V) in human PrP prevents prion disease, however the structural basis for its protective effect remains unknown. Here we show that the mutation alters and constrains the PrP backbone conformation preceding the PrP β-sheet, stabilising PrP dimer interactions by increasing intermolecular hydrogen bonding. It also markedly changes the solution dynamics of the β2-α2 loop, a region of PrP structure implicated in prion transmission and cross-species susceptibility. Both of these structural changes may affect access to protein conformers susceptible to prion formation and explain its profound effect on prion disease.

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Copper and prion diseases

Biochemical Society Transactions ◽

10.1042/bst0300742 ◽

2002 ◽

Vol 30 (4) ◽

pp. 742-745 ◽

Cited By ~ 16

Author(s):

D. R. Brown

Keyword(s):

Prion Protein ◽

Strong Evidence ◽

Prion Diseases ◽

Transmissible Spongiform Encephalopathies ◽

Normal Brain ◽

Spongiform Encephalopathies ◽

Metal Metabolism ◽

Jakob Disease ◽

Anti Oxidant ◽

Experimental Mouse

Transmissible spongiform encephalopathies are diseases of animals and humans that are also termed prion diseases. These diseases are linked together because a normal brain glycoprotein termed the prion protein is converted to a readily detectable protease-resistant isoform. There is now strong evidence to suggest that apart from this difference in resistance a major difference between the isoforms is that the normal prion protein binds copper and has an anti-oxidant function. Brains from Creutzfeldt-Jakob disease patients and brains from mice with experimental mouse scrapie have been shown to have changes in the levels of both copper and manganese. There is growing evidence that links prion diseases to disturbances of metal metabolism.

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Cryo-EM structure of disease-related prion fibrils provides insights into seeding barriers

10.1101/2021.08.10.455830 ◽

2021 ◽

Author(s):

Qiuye Li ◽

Christopher P. Jaroniec ◽

Witold K. Surewicz

Keyword(s):

High Resolution ◽

Prion Protein ◽

Prion Disease ◽

Amyloid Fibrils ◽

Prion Diseases ◽

Protein Aggregates ◽

Direct Support ◽

Human Prion Protein ◽

Structural Insight

One of the least understood aspects of prion diseases is the structure of infectious prion protein aggregates. Here we report a high-resolution cryo-EM structure of amyloid fibrils formed by human prion protein with Y145Stop mutation that is associated with a familial prion disease. This structural insight allows us not only to explain previous biochemical findings, but also provides direct support for the conformational adaptability model of prion transmissibility barriers.

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Prion peptide-mediated calcium level alteration governs neuronal cell damage through AMPK-autophagy flux

10.21203/rs.2.24353/v2 ◽

2020 ◽

Author(s):

Ji-Hong Moon ◽

Sang-Youel Park

Keyword(s):

Cell Death ◽

Intracellular Calcium ◽

Prion Protein ◽

Prion Diseases ◽

Critical Role ◽

Neuronal Cell ◽

Autophagic Cell Death ◽

Autophagy Flux ◽

Human Prion Protein ◽

Ampk Activity

Abstract Background The distinctive molecular structure of the prion protein, PrPsc, is established only in mammals with infectious prion diseases. Prion protein characterizes either the transmissible pathogen itself or a primary constituent of the disease. Our report suggested that prion-mediated neuronal cell death is triggered by the autophagy flux. However, the alteration of intracellular calcium levels, AMPK activity in prion models has not been described. This study is focused on the effect of the changes in intracellular calcium levels on AMPK/autophagy flux pathway and PrP (106–126)-induced neurotoxicity. Methods Western blot and Immunocytochemistry was used to detect AMPK and autophagy-related protein expression. Flow cytometry and a TdT-mediated biotin-16-dUTP nick-end labeling (TUNEL) assay were used to detect the percentage of apoptotic cells. Calcium measurement was employed using fluo-4 by confocal microscope. Results We examined the effect of calcium homeostasis alterations induced by human prion protein on the autophagy flux in neuronal cells. Treatment with human prion protein increased the intracellular calcium concentration and induced cell death in primary neurons as well as in a neuronal cell line. Using pharmacological inhibitors, we showed that the L-type calcium channel is involved in the cellular entry of calcium ions. Inhibition of calcium uptake prevented autophagic cell death and reduction in AMP-activated protein kinase (AMPK) activity induced by human prion protein. Conclusion Our data demonstrated that prion protein-mediated calcium inflow plays a pivotal role in prion protein-induced autophagic cell death, and reduction in AMPK activity in neurons. Altogether, our results suggest that calcium influx might play a critical role in neurodegenerative diseases, including prion diseases.

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PrP glycoforms are associated in a strain-specific ratio in native PrPSc

Journal of General Virology ◽

10.1099/vir.0.80375-0 ◽

2005 ◽

Vol 86 (9) ◽

pp. 2635-2644 ◽

Cited By ~ 45

Author(s):

Azadeh Khalili-Shirazi ◽

Linda Summers ◽

Jacqueline Linehan ◽

Gary Mallinson ◽

David Anstee ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Prion Protein ◽

Prion Disease ◽

Prion Diseases ◽

Cellular Prion Protein ◽

Normal Brain ◽

Western Blots ◽

Prion Strains ◽

Specific Manner ◽

First Time

Prion diseases involve conversion of host-encoded cellular prion protein (PrPC) to a disease-related isoform (PrPSc). Using recombinant human β-PrP, a panel of monoclonal antibodies was produced that efficiently immunoprecipitated native PrPSc and recognized epitopes between residues 93–105, indicating for the first time that this region is exposed in both human vCJD and mouse RML prions. In contrast, monoclonal antibodies raised to human α-PrP were more efficient in immunoprecipitating PrPC than PrPSc, and some of them could also distinguish between different PrP glycoforms. Using these monoclonal antibodies, the physical association of PrP glycoforms was studied in normal brain and in the brains of humans and mice with prion disease. It was shown that while PrPC glycoforms can be selectively immunoprecipitated, the differentially glycosylated molecules of native PrPSc are closely associated and always immunoprecipitate together. Furthermore, the ratio of glycoforms comprising immunoprecipitated native PrPSc from diverse prion strains was similar to those observed on denaturing Western blots. These studies are consistent with the view that the proportion of each glycoform incorporated into PrPSc is probably controlled in a strain-specific manner and that each PrPSc particle contains a mixture of glycoforms.

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Codon 129 polymorphism of the human prion protein influences the kinetics of amyloid formation

Journal of General Virology ◽

10.1099/vir.0.81630-0 ◽

2006 ◽

Vol 87 (8) ◽

pp. 2443-2449 ◽

Cited By ~ 23

Author(s):

Patrick A. Lewis ◽

M. Howard Tattum ◽

Samantha Jones ◽

Daljit Bhelt ◽

Mark Batchelor ◽

...

Keyword(s):

Prion Protein ◽

Amyloid Fibrils ◽

Prion Diseases ◽

Amyloid Formation ◽

Native Structure ◽

Prion Strain ◽

Profound Influence ◽

Human Prion Protein ◽

Kinetics Of ◽

Codon 129

The human prion protein (PrP) has a common polymorphism at residue 129, which can be valine or methionine. This polymorphism has a strong influence on susceptibility to prion diseases and on prion-strain properties. Previous work has shown that this amino acid variation has no measurable effect on the native structure of cellular PrP (PrPC). Here, it is shown that the polymorphism does not change the efficiency of conversion to the β-PrP conformation or affect the binding of copper(II) ions. However, in a partially denatured conformation, the polymorphic variation has a profound influence on the ability of the protein to form amyloid fibrils spontaneously.

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The neuropathological phenotype in transgenic mice expressing different prion protein constructs

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.1994.0038 ◽

1994 ◽

Vol 343 (1306) ◽

pp. 415-423 ◽

Cited By ~ 4

Keyword(s):

Prion Protein ◽

Prion Diseases ◽

Amyloid Plaques ◽

Pathogenic Factor ◽

Wild Type ◽

Vacuolar Degeneration ◽

Neuron Populations ◽

Age Related ◽

Human Prion Protein ◽

The Brain

Neuropathologic examination of transgenic (Tg) mice which express different prion protein (PrP) constructs is essential because spongiform (vacuolar) degeneration of neurons, the distribution of PrP Sc and whether PrP amyloid plaques form are the phenotypes of prion diseases. In Tg models of experimental scrapie, it was found that all of the parameters that define prion isolates (‘strains’) can be manipulated by changing the structure of PrP. In those studies, further evidence that PrP Sc causes scrapie neuropathology and determines scrapie incubation time was obtained. In addition, the distribution of PrP Sc in the brain was unique for each prion isolate. The implications of these findings are first, that prion isolates target different neuron populations for synthesis of nascent pathogenic PrP Sc and, secondly, that prion isolate diversity is determined by neurons. In Tg mice which express mutated PrP mimicking human prion protein genemutations linked to familial prion diseases, the neuropathological changes have been faithfully reproduced. A new age-related, neuromascular disorder has also been identified in uninfected Tg mice which overexpress wild-type PrP C . All of the findings with different PrP constructs plus the absence of scrapie pathology in PrP null mice are the strongest argument that the prion protein is the main etiologic and pathogenic factor of prion disorders.

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Familial human prion diseases associated with prion protein mutations Y226X and G131V are transmissible to transgenic mice expressing human prion protein

Acta Neuropathologica Communications ◽

10.1186/s40478-018-0516-2 ◽

2018 ◽

Vol 6 (1) ◽

Cited By ~ 10

Author(s):

Brent Race ◽

Katie Williams ◽

Andrew G. Hughson ◽

Casper Jansen ◽

Piero Parchi ◽

...

Keyword(s):

Transgenic Mice ◽

Prion Protein ◽

Prion Diseases ◽

Human Prion Protein ◽

Protein Mutations

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

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/gmz124 ◽

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.

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