Abstract
Across the spectrum of sporadic human prion diseases (also known as transmissible spongiform encephalopathies: TSE), there is considerable phenotypic diversity. Cumulative scientific evidence supports that prions, the infectious agents of prion diseases, are constituted predominantly, if not exclusively, by misfolded, typically protease-resistant, disease-associated isoforms of the prion protein (PrPres). Consequently, tissue deposition of PrPres is considered a hallmark of prion disease pathology, and this can be visualized by Western blotting after tissue homogenization and treatment with proteinases, particularly proteinase K (PK). Indeed, Western blot profiles of PrPres are utilized as one marker of different prion strains, with such strains thought to contribute to at least part of the phenotypic variation observed in sporadic human prion disease. Typically, Western blotting of PrPres demonstrates three bands of different electrophoretic mobility, depicting the di-glycosylated, mono-glycosylated and unglycosylated species although further subclassification and the delineation of novel sporadic disease subtypes, such as variably protease-sensitive prionopathy, has contributed greater complexity. Nevertheless, it is the mobility of the unglycosylated PrPres band, the relative abundance of the two glycosylated bands or overall profile of the banding post-PK, in combination with the prion protein gene (PRNP) codon 129 genotype that allows the categorisation of molecular subtypes of sporadic human prion disease. These subtypes appear to correlate with distinct clinico-pathological profiles of sporadic Creutzfeldt-Jakob disease.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Collins S, Boyd A, Lee JS et al (2002) Creutzfeldt-Jakob disease in Australia 1970–1999. Neurology 59:1365–1715
Hill AF, Joiner S, Wadsworth JD et al (2003) Molecular classification of sporadic Creutzfeldt-Jakob disease. Brain 126:1333–1346
Lewis V, Hill AF, Klug GM et al (2005) Australian sporadic CJD analysis supports endogenous determinants of molecular-clinical profiles. Neurology 65:113–118
Parchi P, Giese A, Capellari S et al (1999) Classification of sporadic Creutzfeldt-Jakob disease based on molecular and phenotypic analysis of 300 subjects. Ann Neurol 46:224–233
Zou WQ, Puoti G, Xiao X et al (2010) Variably protease-sensitive prionopathy: a new sporadic disease of the prion protein. Ann Neurol 68:162–172
Gambetti P, Dong Z, Yuan J et al (2008) A novel human disease with abnormal prion protein sensitive to protease. Ann Neurol 63:677–708
Acknowledgements
The Australian National Creutzfeldt-Jakob Disease Registry (ANCJDR) is funded by the Commonwealth Department of Health. The authors thank the families and clinicians for their ongoing support in the surveillance of CJD in Australia. Original T1–T4 standards were supplied to the ANCJDR by the National Institute for Biological Standards and Control (NIBSC) as part of the World Health Organisation Collaborative Nomenclature Study (2001–2002). Since then Australian cases of T1-3 glycotypes have been determined and are used as control samples for future glycotyping gels.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Klug, G.M., Lewis, V., Collins, S.J. (2017). Molecular Subtyping of PrPres in Human Sporadic CJD Brain Tissue. In: Lawson, V. (eds) Prions. Methods in Molecular Biology, vol 1658. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7244-9_23
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7244-9_23
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7242-5
Online ISBN: 978-1-4939-7244-9
eBook Packages: Springer Protocols