Immunopurification of pathological prion protein aggregates

Emiliano Biasini, Laura Tapella, Susanna Mantovani, Matteo Stravalaci, Marco Gobbi, David A. Harris, Roberto Chiesa

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Background: Prion diseases are fatal neurodegenerative disorders that can arise sporadically, be genetically inherited or acquired through infection. The key event in these diseases is misfolding of the cellular prion protein (PrPC) into a pathogenic isoform that is rich in β-sheet structure. This conformational change may result in the formation of PrPSc, the prion isoform of PrP, which propagates itself by imprinting its aberrant conformation onto PrPC molecules. A great deal of effort has been devoted to developing protocols for purifying PrPSc for structural studies, and testing its biological properties. Most procedures rely on protease digestion, allowing efficient purification of PrP27-30, the protease-resistant core of PrPSc. However, protease treatment cannot be used to isolate abnormal forms of PrP lacking conventional protease resistance, such as those found in several genetic and atypical sporadic cases. Principal Findings: We developed a method for purifying pathological PrP molecules based on sequential centrifugation and immunoprecipitation with a monoclonal antibody selective for aggregated PrP. With this procedure we purified full-length PrPSc and mutant PrP aggregates at electrophoretic homogeneity. PrPSc purified from prion-infected mice was able to seed misfolding of PrPC in a protein misfolding cyclic amplification reaction, and mutant PrP aggregates from transgenic mice were toxic to cultured neurons. Significance: The immunopurification protocol described here isolates biologically active forms of aggregated PrP. These preparations may be useful for investigating the structural and chemico-physical properties of infectious and neurotoxic PrP aggregates.

Original languageEnglish
Article numbere7816
JournalPLoS One
Volume4
Issue number11
DOIs
Publication statusPublished - Nov 12 2009

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PrPSc proteins
prions
protein aggregates
Prions
Peptide Hydrolases
proteinases
PrPC Proteins
Protein Isoforms
Prion Diseases
Molecules
Centrifugation
Poisons
mutants
Immunoprecipitation
genomic imprinting
Neurodegenerative Diseases
prion diseases
Transgenic Mice
Neurons
Purification

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Biasini, E., Tapella, L., Mantovani, S., Stravalaci, M., Gobbi, M., Harris, D. A., & Chiesa, R. (2009). Immunopurification of pathological prion protein aggregates. PLoS One, 4(11), [e7816]. https://doi.org/10.1371/journal.pone.0007816

Immunopurification of pathological prion protein aggregates. / Biasini, Emiliano; Tapella, Laura; Mantovani, Susanna; Stravalaci, Matteo; Gobbi, Marco; Harris, David A.; Chiesa, Roberto.

In: PLoS One, Vol. 4, No. 11, e7816, 12.11.2009.

Research output: Contribution to journalArticle

Biasini, E, Tapella, L, Mantovani, S, Stravalaci, M, Gobbi, M, Harris, DA & Chiesa, R 2009, 'Immunopurification of pathological prion protein aggregates', PLoS One, vol. 4, no. 11, e7816. https://doi.org/10.1371/journal.pone.0007816
Biasini E, Tapella L, Mantovani S, Stravalaci M, Gobbi M, Harris DA et al. Immunopurification of pathological prion protein aggregates. PLoS One. 2009 Nov 12;4(11). e7816. https://doi.org/10.1371/journal.pone.0007816
Biasini, Emiliano ; Tapella, Laura ; Mantovani, Susanna ; Stravalaci, Matteo ; Gobbi, Marco ; Harris, David A. ; Chiesa, Roberto. / Immunopurification of pathological prion protein aggregates. In: PLoS One. 2009 ; Vol. 4, No. 11.
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