Diagnosing prion diseases: needs, challenges and hopes

Soto, Claudio

doi:10.1038/nrmicro1003

Cited by 82 publications

(92 citation statements)

References 116 publications

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“…A first possibility is that the association of PrP with raft regions, maintaining the native secondary structure, may exert a protective effect on the protein after synthesis and delivery to the plasma membrane. Moreover, the protein unfolding and increase of oligomerization observed at acidic pH upon interaction with non-raft membranes may be viewed as an early step of the conversion to the aberrant pathogenic form [1], that could be triggered by interaction with endogenously formed or exogenously introduced PrPsc [27], or with chaperone molecules [28].…”

Section: Discussionmentioning

confidence: 99%

Prion protein structure is affected by pH‐dependent interaction with membranes: A study in a model system

Sesana¹,

Barbiroli²,

Bonomi³

et al. 2007

FEBS Letters

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Interaction of full length recombinant hamster prion protein with liposomes mimicking lipid rafts or non-raft membrane regions was studied by circular dichroism, chemical cross-linking and sucrose gradient ultracentrifugation. At pH 7.0, the protein bound palmitoyloleoylphosphatidylcholine/ cholesterol/sphingomyelin/monosialoganglioside GM1 (GM1) ganglioside liposomes but not palmitoyloleoylphosphatidylcholine alone (bound/free = 0.33 and 0.01, respectively), maintaining the native a-helical structure and monomeric form. At pH 5.0, though still binding to quaternary mixtures, in particular GM1, the protein bound also to palmitoyloleoylphosphatidylcholine (bound/free 0.35) becoming unfolded and oligomeric. The pH-dependent interaction with raft or non-raft membranes might have implication in vivo, by stabilizing or destabilizing the protein.

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Section: Discussionmentioning

confidence: 99%

Prion protein structure is affected by pH‐dependent interaction with membranes: A study in a model system

Sesana¹,

Barbiroli²,

Bonomi³

et al. 2007

FEBS Letters

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“…In mammals, a prion (PrP Sc ) is able to facilitate the conversion of the host's normal cellular protein (PrP C ) into PrP Sc and thereby propagate an infection. PrP Sc is believed to be the cause of a group of fatal neurodegenerative diseases called transmissible spongiform encephalopathies (TSEs), known to affect humans and a variety of other animals [2][3][4][5]. Detailed studies show that the normal cellular isoform (PrP C ) and the infectious isoform (PrP Sc ) have identical amino acid sequences and covalent post-translational modifications and, thus, the difference between PrP C and PrP Sc is believed to be exclusively conformational [6 -10].…”

mentioning

confidence: 99%

Mass spectrometric detection of attomole amounts of the prion protein by nanoLC/MS/MS

Onisko¹,

Dynin²,

Requena

et al. 2007

J. Am. Soc. Mass Spectrom.

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Sensitive quantitation of prions in biological samples is an extremely important and challenging analytical problem. Prions are the cause of several fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs). At this time, there are no methods to diagnose TSEs in live animals or to assure a prion-free blood supply for humans. Prions have been shown to be present in blood by transfusion experiments, but based on the amount of infectivity found in these types of experiments, the amount of misfolded prion protein in blood is estimated to be only 30 to 625 amol/mL. More sensitive detection of prions in brain would allow earlier detection of disease and assure a safer food supply. We studied quantitation of the prion protein by use of nanoscale liquid chromatography coupled to a tandem mass spectrometer using the multiple reaction monitoring mode of operation. We developed a method based on the detection of VVEQMCTTQYQK obtained by reduction, alkylation, and digestion with trypsin of the prion protein.

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“…An important effect of this conformational change is that while the entire PrP C is protease sensitive, the C-terminal domain of PrP Sc becomes relatively protease resistant (12,25). Consequently, the presence of protease-resistant PrP Sc in central nervous system (CNS) tissue has provided the basis for the in vitro diagnosis of all prion diseases (19,39). However, available tests are mostly postmortem, invasive, not very sensitive, and nonquantitative.…”

mentioning

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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Diagnosing prion diseases: needs, challenges and hopes

Cited by 82 publications

References 116 publications

Prion protein structure is affected by pH‐dependent interaction with membranes: A study in a model system

Prion protein structure is affected by pH‐dependent interaction with membranes: A study in a model system

Mass spectrometric detection of attomole amounts of the prion protein by nanoLC/MS/MS

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

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