Prion protein expression in different species: Analysis with a panel of new mAbs

Zanusso, Gianluigi; Liu, Dacai; Ferrari, Sergio; Hegyi, Ivan; Yin, Xiaoju; Aguzzi, Adriano; Hornemann, Simone; Liemann, Susanne; Glockshuber, Rudi; Manson, Jean; Brown, Paul; Petersen, Robert B.; Sy, Man Sun

doi:10.1073/pnas.95.15.8812

Cited by 185 publications

(181 citation statements)

References 27 publications

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“…Anti-PrP monoclonal antibody 3F4 (specific to PrP residues 109 -112) was from Richard Kascsak (New York State Institute for Basic Research in Developmental Disabilities). Anti-PrP monoclonal antibody 8H4, which binds to a site between residues 145-180 of PrP, was raised in our facility (20). All other chemicals, including antibodies to the glial fibrillary acidic protein (anti-GFAP) and neurofilament-specific (NF68) antibodies were purchased from Sigma.…”

Section: Methodsmentioning

confidence: 99%

Prion Peptide 106–126 Modulates the Aggregation of Cellular Prion Protein and Induces the Synthesis of Potentially Neurotoxic Transmembrane PrP

Fujioka

Mishra

et al. 2002

Journal of Biological Chemistry

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In infectious and familial prion disorders, neurodegeneration is often seen without obvious deposits of the scrapie prion protein (PrP Sc ), the principal cause of neuronal death in prion disorders. In such cases, neurotoxicity must be mediated by alternative pathways of cell death. One such pathway is through a transmembrane form of PrP. We have investigated the relationship between intracellular accumulation of prion protein aggregates and the consequent up-regulation of transmembrane prion protein in a cell model. Here, we report that exposure of neuroblastoma cells to the prion peptide 106 -126 catalyzes the aggregation of cellular prion protein to a weakly proteinase K-resistant form and induces the synthesis of transmembrane prion protein, the proposed mediator of neurotoxicity in certain prion disorders. The N terminus of newly synthesized transmembrane prion protein is cleaved spontaneously on the cytosolic face of the endoplasmic reticulum, and the truncated C-terminal fragment accumulates on the cell surface. Our results suggest that neurotoxicity in prion disorders is mediated by a complex pathway involving transmembrane prion protein and not by deposits of aggregated and proteinase K-resistant PrP alone.

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Section: Methodsmentioning

confidence: 99%

Prion Peptide 106–126 Modulates the Aggregation of Cellular Prion Protein and Induces the Synthesis of Potentially Neurotoxic Transmembrane PrP

Fujioka

Mishra

et al. 2002

Journal of Biological Chemistry

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“…The generation of recombinant PrP C proteins and anti-PrP monoclonal antibodies (MAbs) was described in detail previously (20,49,50). All MAbs were affinity purified.…”

Section: Methodsmentioning

confidence: 99%

Test for Detection of Disease-Associated Prion Aggregate in the Blood of Infected but Asymptomatic Animals

Chang

Cheng

Yin

et al. 2007

Clin Vaccine Immunol

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We have developed a sensitive in vitro assay for detecting disease-associated prion aggregates by combining an aggregation-specific enzyme-linked immunosorbent assay (AS-ELISA) with the fluorescent amplification catalyzed by T7 RNA polymerase technique (FACTT). The new assay, named aggregation-specific FACTT (AS-FACTT), is much more sensitive than AS-ELISA and could detect prion aggregates in the brain of mice as early as 7 days after an intraperitoneal inoculation of PrP Sc . However, AS-FACTT was still unable to detect prion aggregates in blood of infected mice. To further improve the detection limit of AS-FACTT, we added an additional prion amplification step (Am) and developed a third-generation assay, termed Am-A-FACTT. Am-A-FACTT has 100% sensitivity and specificity in detecting disease-associated prion aggregates in blood of infected mice at late but still asymptomatic stages of disease. At a very early stage, Am-A-FACTT had a sensitivity of 50% and a specificity of 100%. Most importantly, Am-A-FACTT also detects prion aggregates in blood of mule deer infected with the agent causing a naturally occurring prion disease, chronic wasting disease. Application of this assay to cattle, sheep, and humans could safeguard food supplies and prevent human contagion.

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“…Mouse monoclonal antibody 8H4, epitope (residues 145-220) binding is independent of the N-linked glycosylation, because it reacts with both recombinant PrP and all native glycoforms (29). 7A12 (epitope 90 -140) is a mouse monoclonal anti-PrP antibody (30).…”

Section: Antibodiesmentioning

confidence: 99%

Altered Glycosylated PrP Proteins Can Have Different Neuronal Trafficking in Brain but Do Not Acquire Scrapie-like Properties

Cancellotti

Wiseman

Tuzi

et al. 2005

Journal of Biological Chemistry

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N-Linked glycans have been shown to have an important role in the cell biology of a variety of cell surface glycoproteins, including PrP protein. It has been suggested that glycosylation of PrP can influence the susceptibility to transmissible spongiform encephalopathy and determine the characteristics of the many different strains observed in this particular type of disease. To understand the role of carbohydrates in influencing the PrP maturation, stability, and cell biology, we have produced and analyzed gene-targeted murine models expressing differentially glycosylated PrP. Transgenic mice carrying the PrP substitution threonine for asparagine 180 (G1) or threonine for asparagine 196 (G2) or both mutations combined (G3), which eliminate the first, second, and both glycosylation sites, respectively, have been generated by double replacement gene targeting. An in vivo analysis of altered PrP has been carried out in transgenic mouse brains, and our data show that the lack of glycans does not influence PrP maturation and stability. The presence of one chain of sugar is sufficient for the trafficking to the cell membrane, whereas the unglycosylated PrP localization is mainly intracellular. However, this altered cellular localization of PrP does not lead to any overt phenotype in the G3 transgenic mice. Most importantly, we found that, in vivo, unglycosylated PrP does not acquire the characteristics of the aberrant pathogenic form (PrP Sc ), as was previously reported using in vitro models.

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Prion protein expression in different species: Analysis with a panel of new mAbs

Cited by 185 publications

References 27 publications

Prion Peptide 106–126 Modulates the Aggregation of Cellular Prion Protein and Induces the Synthesis of Potentially Neurotoxic Transmembrane PrP

Prion Peptide 106–126 Modulates the Aggregation of Cellular Prion Protein and Induces the Synthesis of Potentially Neurotoxic Transmembrane PrP

Test for Detection of Disease-Associated Prion Aggregate in the Blood of Infected but Asymptomatic Animals

Altered Glycosylated PrP Proteins Can Have Different Neuronal Trafficking in Brain but Do Not Acquire Scrapie-like Properties

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